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Subject: Oncomirs [mdash] microRNAs with a role in cancer : Article : Nature Reviews Cancer
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<H1 class=3Dpage-header>Review</H1></DIV>
<DIV class=3Dbox>
<P id=3Dcite><SPAN class=3Djournalname>Nature Reviews Cancer</SPAN> =
<SPAN=20
class=3Djournalnumber>6</SPAN>, <SPAN class=3Dcite-pages>259-269</SPAN> =
<SPAN=20
class=3Dcite-month-year>(April 2006)</SPAN><SPAN class=3Ddivider> | =
</SPAN><SPAN=20
class=3Dcite-doi><SPAN class=3Ddoi><ABBR=20
title=3D"Digital Object =
Identifier">doi</ABBR>:10.1038/nrc1840</SPAN></SPAN></P>
<P id=3Derrorcor></P>
<H2 id=3Datl>Oncomirs =97 microRNAs with a role in cancer</H2>
<P id=3Daug>Aurora Esquela-Kerscher<SUP><A title=3D"affiliated with "=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#a1">1</=
A></SUP>=20
&amp; Frank J. Slack<SUP><A title=3D"affiliated with "=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#a1">1</=
A></SUP>&nbsp;=20
<A title=3D"More information about the author"=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/authors/nrc1840.html">Abo=
ut the=20
authors</A></P>
<DIV id=3Dabs><A class=3Dbacktotop=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#top">To=
p<SPAN=20
class=3Dhidden> of page</SPAN></A>
<H3 class=3Dhidden>Abstract</H3>
<P class=3Dlead>MicroRNAs (miRNAs) are an abundant class of small=20
non-protein-coding RNAs that function as negative gene regulators. They =
regulate=20
diverse biological processes, and bioinformatic data indicates that each =
miRNA=20
can control hundreds of gene targets, underscoring the potential =
influence of=20
miRNAs on almost every genetic pathway. Recent evidence has shown that =
miRNA=20
mutations or mis-expression correlate with various human cancers and =
indicates=20
that miRNAs can function as tumour suppressors and oncogenes. miRNAs =
have been=20
shown to repress the expression of important cancer-related genes and =
might=20
prove useful in the diagnosis and treatment of cancer.</P></DIV>
<UL class=3Dswitch-article-view>
  <LI>View <A title=3D""=20
  =
href=3D"http://www.nature.com/nrc/journal/v6/n4/execsumm/nrc1840.html">At=
 a=20
  Glance</A></LI></UL>
<DIV id=3Darticlebody>
<P class=3Dnorm>Cancer is caused by uncontrolled proliferation and the=20
inappropriate survival of damaged cells, which results in tumour =
formation.=20
Cells have developed several safeguards to ensure that cell division,=20
differentiation and death occur correctly and in a coordinated fashion, =
both=20
during development and in the adult body. Many regulatory factors switch =
on or=20
off genes that direct cellular proliferation and differentiation. Damage =
to=20
these genes, which are referred to as tumour-suppressor genes and =
oncogenes, is=20
selected for in cancer. Most tumour-suppressor genes and oncogenes are =
first=20
transcribed from DNA into RNA, and are then translated into protein to =
exert=20
their effects. Recent evidence indicates that small non-protein-coding =
RNA=20
molecules, called microRNAs (miRNAs), might also function as tumour =
suppressors=20
and oncogenes.</P>
<P class=3Dnorm>We are just beginning to understand how this novel class =
of gene=20
regulators is involved in cancer-related pro-cesses in humans. Out of =
hundreds=20
of miRNAs that have recently been identified, only a small number from =
worm, fly=20
and human genomes have been characterized. They have been shown to =
control cell=20
growth, differentiation and apoptosis; consequently, impaired miRNA =
expression=20
has been implicated in tumorigenesis. Continued research into miRNA =
function=20
might lead to an advanced understanding of the mechanisms that lead to=20
tumorigenesis. In this Review, we discuss the emerging field of =
'oncomirs' =97=20
miRNAs that are associated with cancer =97 and how these miRNAs could be =
used in=20
the diagnosis and treatment of cancer.</P>
<H4 class=3Dnorm =
id=3DmiRNAs-are-a-large-family-of-gene-regulators>miRNAs are a=20
large family of gene regulators</H4>
<P class=3Dnorm>miRNAs are non-coding, single-stranded RNAs of <IMG=20
style=3D"BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; VERTICAL-ALIGN: middle; BORDER-RIGHT-WIDTH: =
0px"=20
alt=3Dapprox=20
src=3D"http://www.nature.com/__chars/math/special/sim/black/med/base/glyp=
h.gif">22=20
nucleotides and constitute a novel class of gene regulators that are =
found in=20
both plants and animals<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B1">1</=
A></SUP>.=20
They negatively regulate their targets in one of two ways depending on =
the=20
degree of complementarity between the miRNA and the target (<A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#f1">Fig=
. 1</A>).=20
First, miRNAs that bind with perfect or nearly perfect complementarity =
to=20
protein-coding mRNA sequences induce the RNA-mediated interference =
(RNAi)=20
pathway (see Ref. <A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B2">2</=
A> for a=20
review on RNAi). Briefly, mRNA transcripts are cleaved by ribonucleases =
in the=20
miRNA-associated, multiprotein RNA-induced-silencing complex (miRISC), =
which=20
results in the degradation of target mRNAs. This mechanism of =
miRNA-mediated=20
gene silencing is commonly found in plants<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B3">3, =

</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B4">4, =

</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B5">5</=
A></SUP>,=20
but miRNA-directed mRNA cleavage has also been shown to occur in =
mammals<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B6">6</=
A></SUP>.=20
However, most animal miRNAs are thought to use a second mechanism of =
gene=20
regulation that does not involve the cleavage of their mRNA targets. =
These=20
miRNAs exert their regulatory effects by binding to imperfect =
complementary=20
sites within the 3' untranslated regions (UTRs) of their mRNA targets, =
and they=20
repress target-gene expression post-transcriptionally, apparently at the =
level=20
of translation, through a RISC complex that is similar to, or possibly =
identical=20
with, the one that is used for the RNAi pathway<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B7">7, =

</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B8">8, =

</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B9">9, =

</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B10">10=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B11">11=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B12">12=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B13">13=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B14">14=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B15">15=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B16">16=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B17">17=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B18">18=
</A></SUP>.=20
Consistent with translational control, miRNAs that use this mechanism =
reduce the=20
protein levels of their target genes, but the mRNA levels of these genes =
are=20
barely affected. However, recent findings indicate that miRNAs that =
share only=20
partial complementarity with their targets can also induce mRNA =
degradation, but=20
it is unclear if translational inhibition precedes destabilization of =
the gene=20
targets in these cases<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B19">19=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B20">20=
</A></SUP>.</P>
<DIV class=3Dfigure-table id=3Df1>
<H5 class=3Dnorm><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_F1.html">=
Figure=20
1<SPAN class=3Ddivider> | </SPAN>The biogenesis of microRNAs.</A></H5><A =

href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_F1.html">=
<IMG=20
class=3Dthumb=20
alt=3D"Figure 1 : The biogenesis of microRNAs. Unfortunately we are =
unable to provide accessible alternative text for this. If you require =
assistance to access this image, or to obtain a text description, please =
contact npg@nature.com"=20
src=3D"http://www.nature.com/nrc/journal/v6/n4/thumbs/nrc1840-f1.jpg"></A=
>
<P>MicroRNA (miRNA) genes are generally transcribed by RNA Polymerase II =
(Pol=20
II) in the nucleus to form large pri-miRNA transcripts, which are capped =

(<SUP>7</SUP>MGpppG) and polyadenylated (AAAAA). These pri-miRNA =
transcripts are=20
processed by the RNase III enzyme Drosha and its co-factor, Pasha, to =
release=20
the <IMG=20
style=3D"BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; VERTICAL-ALIGN: middle; BORDER-RIGHT-WIDTH: =
0px"=20
alt=3Dapprox=20
src=3D"http://www.nature.com/__chars/math/special/sim/black/med/base/glyp=
h.gif">70-nucleotide=20
pre-miRNA precursor product. (Note that the human <SPAN =
class=3Di>let-7a-1</SPAN>=20
miRNA is shown here as an example of a pre-miRNA hairpin sequence. The =
mature=20
miRNA sequence is shown in red.) RAN=96GTP and exportin 5 transport the =
pre-miRNA=20
into the cytoplasm. Subsequently, another RNase III enzyme, Dicer, =
processes the=20
pre-miRNA to generate a transient <IMG=20
style=3D"BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; VERTICAL-ALIGN: middle; BORDER-RIGHT-WIDTH: =
0px"=20
alt=3Dapprox=20
src=3D"http://www.nature.com/__chars/math/special/sim/black/med/base/glyp=
h.gif">22-nucleotide=20
miRNA:miRNA<SUP>*</SUP> duplex. This duplex is then loaded into the=20
miRNA-associated multiprotein RNA-induced silencing complex (miRISC) =
(light=20
blue), which includes the Argonaute proteins, and the mature =
single-stranded=20
miRNA (red) is preferentially retained in this complex. The mature miRNA =
then=20
binds to complementary sites in the mRNA target to negatively regulate =
gene=20
expression in one of two ways that depend on the degree of =
complementarity=20
between the miRNA and its target. miRNAs that bind to mRNA targets with=20
imperfect complementarity block target gene expression at the level of =
protein=20
translation (lower left). However, recent evidence indicates that miRNAs =
might=20
also affect mRNA stability (not shown). Complementary sites for miRNAs =
using=20
this mechanism are generally found in the 3' untranslated regions (3' =
UTRs) of=20
the target mRNA genes. miRNAs that bind to their mRNA targets with =
perfect (or=20
nearly perfect) complementarity induce target-mRNA cleavage (lower =
right).=20
miRNAs using this mechanism bind to miRNA complementary sites that are =
generally=20
found in the coding sequence or open reading frame (ORF) of the mRNA =
target.</P>
<UL class=3Doptions>
  <LI class=3Dhigh-res><A=20
  =
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_F1.html">=
High=20
  resolution image and legend (42 <ABBR title=3DKilobyte>KB</ABBR>) </A>
  <LI class=3Dfti><A title=3D"View a list of all figures and tables in =
this article"=20
  =
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_ft.html">=
Figures=20
  and tables index</A>
  <LI class=3Dpowerpoint><A class=3Ddownload-pp=20
  =
href=3D"http://www.nature.com/nrc/journal/v6/n4/slides/nrc1840-pf1.ppt">D=
ownload=20
  Power<SPAN class=3Dhidden> </SPAN>Point slide (250 <ABBR=20
  title=3DKilobyte>KB</ABBR>) </A></LI></UL><SPAN=20
class=3Dcleardiv><!-- --></SPAN></DIV><BR class=3Dclear>
<P class=3Dnorm>The biogenesis of miRNAs has only recently been =
elucidated<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B1">1</=
A></SUP>=20
(<A =
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#f1">Fig=
.=20
1</A>). miRNAs, which generally seem to be transcribed by RNA polymerase =
II, are=20
initially made as large RNA precursors that are called pri-miRNAs. The=20
pri-miRNAs are processed in the nucleus by the RNase III enzyme, <A=20
href=3D"http://us.expasy.org/uniprot/Q9NRR4">Drosha</A>, and the=20
double-stranded-RNA-binding protein, <A=20
href=3D"http://us.expasy.org/uniprot/Q8WYQ5">Pasha</A> (also known as =
DGCR8), into=20
<IMG=20
style=3D"BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; VERTICAL-ALIGN: middle; BORDER-RIGHT-WIDTH: =
0px"=20
alt=3Dapprox=20
src=3D"http://www.nature.com/__chars/math/special/sim/black/med/base/glyp=
h.gif">70-nucleotide=20
pre-miRNAs, which fold into imperfect stem-loop structures<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B21">21=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B22">22=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B23">23=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B24">24=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B25">25=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B26">26=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B27">27=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B28">28=
</A></SUP>.=20
The pre-miRNAs are then exported into the cytoplasm by the RAN =
GTP-dependent=20
transporter <A href=3D"http://us.expasy.org/uniprot/Q9ULC9">exportin =
5</A> (Refs=20
<SUP><A =
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B29">29=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B30">30=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B31">31=
</A></SUP>)=20
and undergo an additional processing step in which a double-stranded RNA =
of <IMG=20
style=3D"BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; VERTICAL-ALIGN: middle; BORDER-RIGHT-WIDTH: =
0px"=20
alt=3Dapprox=20
src=3D"http://www.nature.com/__chars/math/special/sim/black/med/base/glyp=
h.gif">22=20
nucleotides in length, referred to as the miRNA:miRNA<SUP>*</SUP> =
duplex, is=20
excised from the pre-miRNA hairpin by another RNAse III enzyme, <A=20
href=3D"http://us.expasy.org/uniprot/Q9UPY3">Dicer</A><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B19">19=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B32">32=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B33">33=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B34">34=
</A></SUP>.=20
Subsequently, the miRNA:miRNA<SUP>*</SUP> duplex is incorporated into =
the miRISC=20
complex. The mature miRNA strand is preferentially retained in the =
functional=20
miRISC complex and negatively regulates its target genes (see =
above).</P>
<P class=3Dnorm>Many researchers failed to appreciate the importance of =
miRNAs=20
when they were initially described more than a decade ago. This is =
because the=20
first miRNA gene that was identified, <SPAN class=3Di>lin-4</SPAN>, was =
thought to=20
be unique to the roundworm <SPAN class=3Di>Caenorhabditis elegans</SPAN> =
and=20
control the timing and progression of the nematode life cycle<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B8">8, =

</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B12">12=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B35">35=
</A></SUP>.=20
However, the identification of hundreds of miRNAs within worm, fly and =
mammalian=20
genomes by traditional cloning techniques and bioinformatics<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B12">12=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B17">17=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B21">21=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B36">36=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B37">37=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B38">38=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B39">39=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B40">40=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B41">41=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B42">42=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B43">43=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B44">44=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B45">45=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B46">46=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B47">47=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B48">48=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B49">49=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B50">50=
</A></SUP>=20
has now captured the attention of scientists specializing in various =
different=20
fields. There are estimated to be at least 300 miRNAs (and there might =
be as=20
many as 1000) in the human genome, comprising <IMG=20
style=3D"BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; VERTICAL-ALIGN: middle; BORDER-RIGHT-WIDTH: =
0px"=20
alt=3Dapprox=20
src=3D"http://www.nature.com/__chars/math/special/sim/black/med/base/glyp=
h.gif">1=964%=20
of all expressed human genes, which makes miRNAs one of the largest =
classes of=20
gene regulator<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B46">46=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B51">51=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B52">52=
</A></SUP>.=20
Most human miRNAs are found within introns of either protein-coding or =
noncoding=20
mRNA transcripts<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B53">53=
</A></SUP>.=20
The remaining miRNAs are either located far from other transcripts in =
the=20
genome, within the exons of noncoding mRNA genes or within the 3' UTRs =
of mRNA=20
genes, or they are clustered with other miRNA genes, including a cluster =
on=20
chromosome 19 that is comprised of 54 novel miRNAs<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B1">1, =

</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B27">27=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B52">52=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B53">53=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B54">54=
</A></SUP>.=20
miRNAs can be grouped into families on the basis of sequence homology, =
which is=20
found primarily at the 5' end of the mature miRNAs, but whether members =
of the=20
same miRNA family control similar biological events remains to be seen. =
Many=20
miRNAs are evolutionarily conserved from worms to humans, which implies =
that=20
these miRNAs direct essential processes both during development and in =
the adult=20
body.</P>
<H4 class=3Dnorm id=3DmiRNAs-regulate-multiple-gene-targets>miRNAs =
regulate multiple=20
gene targets</H4>
<P class=3Dnorm>The current challenge is to accurately identify targets =
that are=20
regulated by miRNAs. Because miRNAs usually bind to their targets with=20
incomplete complementarity, which allows short stretches of mismatched=20
base-pairs and G=96U base pairing, the identification of gene targets =
with a=20
simple BLAST search is impossible. However, current bioinformatic =
approaches=20
have taken advantage of the fact that miRNAs within families have =
highest=20
homology at the 5' end of the mature miRNA. Several studies have =
indicated that=20
the 5' end of the miRNA is crucial for the stability and proper loading =
of the=20
miRNA into the miRISC complex<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B55">55=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B56">56=
</A></SUP>,=20
and this end is also important for biological function<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B57">57=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B58">58=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B59">59=
</A></SUP>.=20
Therefore, most bioinformatic algorithms use a 'miRNA seed' that =
encompasses the=20
first 2=968 bases of the mature miRNA sequence to search for =
complementarity to=20
sequences in the 3' UTR of all expressed genes. These studies have =
revealed that=20
a single miRNA might bind to as many as 200 gene targets and that these =
targets=20
can be diverse in their function; they include transcription factors, =
secreted=20
factors, receptors and transporters<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B20">20=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B47">47=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B60">60=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B61">61=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B62">62=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B63">63=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B64">64=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B65">65=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B66">66=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B67">67=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B68">68=
</A></SUP>.=20
So, miRNAs potentially control the expression of about one-third of =
human=20
mRNAs.</P>
<P class=3Dnorm>However, many targets are missed using the 'miRNA seed' =
approach=20
as there is evidence that other variables besides the first 2=968 =
nucleotides of=20
the miRNA sequence dictate miRNA regulation. For example, the complete =
mature=20
miRNA sequence of <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D3772493"><SP=
AN=20
class=3Di>let-7</SPAN></A> has been evolutionarily conserved from worms =
to=20
humans<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B37">37=
</A></SUP>,=20
demonstrating the functional relevance of the 3' end. Furthermore, =
mutagenesis=20
studies in our laboratory have shown that base pairing at both the 5' =
and 3'=20
ends of <SPAN class=3Di>let-7</SPAN> is required for downregulation of a =
target=20
gene<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B69">69=
</A></SUP>.=20
Therefore, the number of known miRNA targets could increase and the =
challenge=20
will be to determine which predicted targets are biologically relevant. =
Sites=20
that are complementary to several miRNAs have also been identified =
within the 3'=20
UTR of a single gene target, indicating that complex patterns of =
combinatorial=20
regulation by miRNAs exist<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B12">12=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B16">16=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B61">61=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B70">70=
</A></SUP>.=20
Because miRNAs potentially have a broad influence over several diverse =
genetic=20
pathways, the deletion or misexpression of these small RNAs is likely to =
be=20
pleotropic and contribute to disease, including cancer.</P>
<H4 class=3Dnorm id=3DThe-miRNAassociated-genes-and-human-cancer>The=20
miRNA-associated genes and human cancer</H4>
<P class=3Dnorm>Components of the miRNA-machinery have been implicated =
in=20
tumorigenesis. Expression of Dicer has been shown to be downregulated in =
<A=20
href=3D"http://www.cancer.gov/cancertopics/types/lung">lung =
cancer</A><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B71">71=
</A></SUP>.=20
Karube <SPAN class=3Di>et al</SPAN>. examined RNA expression levels of =
<SPAN=20
class=3Di>DICER</SPAN> and <SPAN class=3Di>DROSHA</SPAN> in 67 =
non-small-cell lung=20
cancer samples<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B71">71=
</A></SUP>.=20
They found that reduced expression of <SPAN class=3Di>DICER</SPAN> =
correlates with=20
shortened post-operative survival, which indicates that Dicer might be =
able to=20
prevent the transformation of lung tissue. As Dicer has been implicated =
in=20
heterochromatin maintenance and centromeric silencing, reduced protein =
levels=20
might directly result in genomic instability and lead to tumour =
formation<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B71">71=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B72">72=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B73">73=
</A></SUP>.=20
However, Kanellopoulou <SPAN class=3Di>et al</SPAN>. recently showed =
that mouse=20
embryonic stem (ES) cells that lack <SPAN class=3Di>Dicer</SPAN> showed =
no=20
aberrations in chromosome number or structure despite observed =
heterochromatin=20
deficits, and the injection of these <SPAN =
class=3Di>Dicer</SPAN>-deficient ES=20
cells into nude mice failed to generate tumours<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B72">72=
</A></SUP>.=20
Therefore, the potential role of Dicer in tumour formation might be =
indirect and=20
result from the reduction of miRNAs in the lung that posses =
tumour-suppressor=20
characteristics. <SPAN class=3Di>Dicer</SPAN> gene-disruption studies in =
mice have=20
shown that this gene is important during mammalian development and =
directs=20
proper stem-cell differentiation, T-cell specification and limb =
morphogenesis,=20
presumably reflecting the role of mature miRNAs in these =
processes<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B72">72=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B74">74=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B75">75=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B76">76=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B77">77=
</A></SUP>.=20
Therefore, the association of <SPAN class=3Di>Dicer</SPAN> with lung =
cancer=20
indicates a further role for this gene, and perhaps miRNAs, during lung=20
differentiation.</P>
<P class=3Dnorm>The Argonaute proteins, which are crucial components of =
the RISC=20
complex that direct both short interfering (siRNA)- and miRNA-mediated =
gene=20
regulation, have also been associated with various cancers. Three human=20
Argonaute genes =97 <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D192669"><SPA=
N=20
class=3Di>AGO3</SPAN></A> (also known as eukaryotic =
translation-initiation factor=20
2 subunit 3 (<SPAN class=3Di>EIF2C3</SPAN>)), <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D26523"><SPAN=
=20
class=3Di>AGO1</SPAN></A> (also known as <SPAN class=3Di>EIF2C1</SPAN>) =
and <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D192670"><SPA=
N=20
class=3Di>AGO4</SPAN></A> (also known as <SPAN class=3Di>EIF2C4</SPAN>), =
which are=20
clustered on chromosome 1 (1p34=9635) =97 are frequently deleted in <A=20
href=3D"http://www.cancer.gov/cancertopics/types/wilms">Wilms tumour</A> =
of the=20
kidney and have also been associated with neuro-ectodermal =
tumours<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B7">7, =

</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B78">78=
</A></SUP>.=20
<SPAN class=3Di>AGO1</SPAN> is highly expressed in the developing lung =
and kidney=20
and is notably increased in <A=20
href=3D"http://www.cancer.gov/cancertopics/types/kidney">renal =
tumours</A> that=20
lack the Wilms-tumour suppressor gene, <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D7490">WT1</A=
>=20
(Ref. <A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B7">7</=
A>). This=20
indicates a role for <SPAN class=3Di>AGO1</SPAN> in the differentiation =
of these=20
tissues during embryogenesis. An additional Argonaute gene, <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D9271"><SPAN =

class=3Di>HIWI</SPAN></A>, which is the human orthologue of the <SPAN=20
class=3Di>Drosophila melanogaster</SPAN> Argonaute gene <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D34521"><SPAN=
=20
class=3Di>piwi</SPAN></A>, maps to the genomic region 12q24.33, which is =
a locus=20
that is associated with <A=20
href=3D"http://www.cancer.gov/cancertopics/types/testicular">testicular =
germ-cell=20
cancers</A><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B79">79=
</A></SUP>.=20
This testis-specific gene was found to be upregulated in 12 out of 19 =
testicular=20
seminomas (tumours that are derived from embryonic germ cells). These =
studies=20
indicate that <SPAN class=3Di>HIWI</SPAN> might have oncogenic activity =
and might=20
normally function to control the proliferation and maintenance of germ =
cells.=20
Further insight into the accessory proteins that comprise the machinery=20
mediating miRNA processing and miRNA-directed gene repression will be =
important=20
when designing therapeutic drugs that use miRNAs to treat disorders such =
as=20
cancer.</P>
<H4 class=3Dnorm id=3DmiRNAs-growth-differentiation-and-disease>miRNAs, =
growth,=20
differentiation and disease</H4>
<P class=3Dnorm>The biological roles of only a small fraction of =
identified miRNAs=20
have been elucidated to date. These miRNAs regulate cancer-related =
processes=20
such as cell growth and tissue differentiation, and therefore might =
themselves=20
function as oncomirs. For example, the miRNAs that are encoded by <SPAN=20
class=3Di>lin-4</SPAN> and <SPAN class=3Di>let-7</SPAN> control the =
timing of cell=20
differentiation and proliferation in <SPAN class=3Di>C. elegans</SPAN>. =
Mutations=20
in these <SPAN class=3Di>C. elegans</SPAN> miRNA genes lead to =
abnormalities in=20
cell-cycle exit and terminal differentiation, which are characteristics =
that are=20
found in cancer cells. Interestingly, the mammalian homologues of <SPAN=20
class=3Di>lin-4</SPAN> and <SPAN class=3Di>let-7</SPAN> have been shown =
to control=20
cell proliferation in human cell lines<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B80">80=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B81">81=
</A></SUP>=20
and are also associated with various cancers<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B81">81=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B82">82=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B83">83=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B84">84=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B85">85=
</A></SUP>=20
(discussed below).</P>
<P class=3Dnorm>The <SPAN class=3Di>D. melanogaster</SPAN> miRNA that is =
encoded by=20
<A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D3772549"><SP=
AN=20
class=3Di>bantam</SPAN></A> induces tissue growth by both stimulating =
cell=20
proliferation and inhibiting apoptosis<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B17">17=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B86">86=
</A></SUP>,=20
which are features that are shared with <SPAN class=3Di>bona fide</SPAN> =

oncogenes. However, there are no close homologues of <SPAN =
class=3Di>bantam</SPAN>=20
in humans. Another <SPAN class=3Di>D. melanogaster</SPAN> miRNA, <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D3772293">miR=
-14</A>,=20
is a strong suppressor of apoptosis, which is also an important =
oncogenic=20
characteristic<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B49">49=
</A></SUP>.=20
In addition, miR-14 also seems to have unrelated functions in the <SPAN=20
class=3Di>D. melanogaster</SPAN> stress-response pathway and in =
regulating fat=20
metabolism<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B49">49=
</A></SUP>.=20
Other characterized miRNAs have essential functions during development =
and=20
direct the proper differentiation of cells into various tissues. =
Examples=20
include <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D3565055">miR=
-273</A>=20
and the miRNA that is encoded by <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D178087"><SPA=
N=20
class=3Di>lys-6</SPAN></A>, which are involved in patterning the <SPAN =
class=3Di>C.=20
elegans</SPAN> nervous system<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B43">43=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B50">50=
</A></SUP>;=20
miR-430 in <SPAN class=3Di>Danio rerio</SPAN> brain development<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B87">87=
</A></SUP>;=20
<A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D387176">miR-=
181</A>=20
in the differentiation of mammalian haematopoietic cells towards the =
B-cell=20
lineage<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B88">88=
</A></SUP>;=20
<A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D494324">miR-=
375</A>=20
in mammalian pancreatic islet-cell development and the regulation of =
insulin=20
secretion<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B89">89=
</A></SUP>;=20
<A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406935">miR-=
143</A>=20
during mammalian adipocyte differentiation<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B90">90=
</A></SUP>;=20
miR-196 in mammalian limb patterning<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B91">91=
</A></SUP>;=20
and the miR-1 genes during mammalian heart development<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B92">92=
</A></SUP>.=20
Further characterization of novel miRNAs might reveal other gene =
regulators that=20
coordinate proper organ formation, embryonic patterning and body growth, =
and=20
might also provide insight into the mechanisms of human diseases such as =

cancer.</P>
<H4 class=3Dnorm id=3DmiRNAs-that-are-implicated-in-cancer>miRNAs that =
are=20
implicated in cancer</H4>
<P class=3Dnorm>miRNA expression correlates with various cancers, and =
these genes=20
are thought to function as both tumour suppressors and oncogenes (<A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#f2">Fig=
. 2</A>;=20
<A =
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#t1">Tab=
le=20
1</A>). A recent study showed that about 50% of annotated human miRNAs =
are=20
located in areas of the genome, known as <A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/glossary/nrc1840.html#df1=
">fragile=20
sites</A>, that are associated with cancer. This indicates that miRNAs =
might=20
have a crucial function in cancer progression<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B83">83=
</A></SUP>.=20
For example, <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406911"><SPA=
N=20
class=3Di>mir-125b-1</SPAN></A>, the homologue of <SPAN class=3Di>C. =
elegans=20
lin-4</SPAN>, is located in a fragile site on chromosome 11q24, which is =
deleted=20
in a subset of patients with <A=20
href=3D"http://www.cancer.gov/cancertopics/types/breast">breast</A>, =
lung, <A=20
href=3D"http://www.cancer.gov/cancertopics/types/ovarian">ovarian</A> =
and <A=20
href=3D"http://www.cancer.gov/cancertopics/types/cervical">cervical=20
cancer</A><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B83">83=
</A></SUP>.=20
A report by Sonoki <SPAN class=3Di>et al</SPAN>. also linked this gene =
with <A=20
href=3D"http://www.cancer.gov/cancertopics/types/leukemia">leukaemia</A>,=
 and=20
described a patient with precursor-B-cell acute lympho-blastic leukaemia =
who=20
carried an insertion of pre-miRNA into the immunoglobulin heavy-chain=20
locus<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B85">85=
</A></SUP>.=20
Although the investigators could not determine how <SPAN=20
class=3Di>mir-125b-1</SPAN> expression<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B85">85=
</A></SUP>=20
was modulated in the tumour cells, this study supports a role for this =
gene as=20
an oncomir.</P>
<DIV class=3Dfigure-table id=3Df2>
<H5 class=3Dnorm><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_F2.html">=
Figure=20
2<SPAN class=3Ddivider> | </SPAN>MicroRNAs can function as tumour =
suppressors and=20
oncogenes.</A></H5><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_F2.html">=
<IMG=20
class=3Dthumb=20
alt=3D"Figure 2 : MicroRNAs can function as tumour suppressors and =
oncogenes. Unfortunately we are unable to provide accessible alternative =
text for this. If you require assistance to access this image, or to =
obtain a text description, please contact npg@nature.com"=20
src=3D"http://www.nature.com/nrc/journal/v6/n4/thumbs/nrc1840-f2.jpg"></A=
>
<P><SPAN class=3Db>a</SPAN> | In normal tissues, proper microRNA (miRNA) =

transcription, processing and binding to complementary sequences on the =
target=20
mRNA results in the repression of target-gene expression through a block =
in=20
protein translation or altered mRNA stability (not shown). The overall =
result is=20
normal rates of cellular growth, proliferation, differentiation and cell =
death.=20
<SPAN class=3Db>b</SPAN> | The reduction or deletion of a miRNA that =
functions as=20
a tumour suppressor leads to tumour formation. A reduction in or =
elimination of=20
mature miRNA levels can occur because of defects at any stage of miRNA=20
biogenesis (indicated by question marks) and ultimately leads to the=20
inappropriate expression of the miRNA-target oncoprotein (purple =
squares). The=20
overall outcome might involve increased proliferation, invasiveness or=20
angiogenesis, decreased levels of apoptosis, or undifferentiated or=20
de-differentiated tissue, ultimately leading to tumour formation. <SPAN=20
class=3Db>c</SPAN> | The amplification or overexpression of a miRNA that =
has an=20
oncogenic role would also result in tumour formation. In this situation, =

increased amounts of a miRNA, which might be produced at inappropriate =
times or=20
in the wrong tissues, would eliminate the expression of a miRNA-target=20
tumour-suppressor gene (pink) and lead to cancer progression. Increased =
levels=20
of mature miRNA might occur because of amplification of the miRNA gene, =
a=20
constitutively active promoter, increased efficiency in miRNA processing =
or=20
increased stability of the miRNA (indicated by question marks). ORF, =
open=20
reading frame.</P>
<UL class=3Doptions>
  <LI class=3Dhigh-res><A=20
  =
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_F2.html">=
High=20
  resolution image and legend (64 <ABBR title=3DKilobyte>KB</ABBR>) </A>
  <LI class=3Dfti><A title=3D"View a list of all figures and tables in =
this article"=20
  =
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_ft.html">=
Figures=20
  and tables index</A>
  <LI class=3Dpowerpoint><A class=3Ddownload-pp=20
  =
href=3D"http://www.nature.com/nrc/journal/v6/n4/slides/nrc1840-pf2.ppt">D=
ownload=20
  Power<SPAN class=3Dhidden> </SPAN>Point slide (272 <ABBR=20
  title=3DKilobyte>KB</ABBR>) </A></LI></UL><SPAN=20
class=3Dcleardiv><!-- --></SPAN></DIV><BR class=3Dclear>
<DIV class=3Dfigure-table id=3Dt1>
<H5 class=3Dnorm><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_T1.html">=
Table=20
1<SPAN class=3Ddivider> | </SPAN>MicroRNAs that are associated with =
human=20
cancers</A></H5><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_T1.html">=
<IMG=20
class=3Dthumb alt=3D"Table 1 - MicroRNAs that are associated with human =
cancers"=20
src=3D"http://www.nature.com/common/images/table_thumb.gif"></A>
<UL class=3Doptions>
  <LI class=3Dfull><A=20
  =
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_T1.html">=
Full=20
  table</A>=20
  <LI class=3Dfti><A title=3D"View a list of all figures and tables in =
this article"=20
  =
href=3D"http://www.nature.com/nrc/journal/v6/n4/fig_tab/nrc1840_ft.html">=
Figures=20
  and tables index</A>
  <LI class=3Dpowerpoint><A class=3Ddownload-pp=20
  =
href=3D"http://www.nature.com/nrc/journal/v6/n4/slides/nrc1840-pt1.ppt">D=
ownload=20
  Power<SPAN class=3Dhidden> </SPAN>Point slide (262 <ABBR=20
  title=3DKilobyte>KB</ABBR>) </A></LI></UL><SPAN=20
class=3Dcleardiv><!-- --></SPAN></DIV><BR class=3Dclear>
<P class=3Dnorm>The first indication that miRNAs could function as =
tumour=20
suppressors came from a report by Calin <SPAN class=3Di>et al</SPAN>. =
that showed=20
that patients who were diagnosed with a common form of adult leukaemia, =
B-cell=20
chronic lymphocytic leukemia (CLL), often have deletions or =
downregulation of=20
two clustered miRNA genes, <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406948"><SPA=
N=20
class=3Di>mir-15a</SPAN></A> and <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406950"><SPA=
N=20
class=3Di>mir-16-1</SPAN></A> (Ref. <A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B93">93=
</A>).=20
Deletions within the 13q14 locus occur in more than 65% of CLL cases, as =
well as=20
in 50% of mantle cell lymphomas, 16=9640% of <A=20
href=3D"http://www.cancer.gov/cancertopics/types/myeloma">multiple =
myelomas</A>=20
and 60% of <A =
href=3D"http://www.cancer.gov/cancertopics/types/prostate">prostate=20
cancers</A>. Therefore, it was predicted that a tumour-suppressor gene =
must=20
reside in this 30-kb region. Interestingly, <SPAN =
class=3Di>mir-15a</SPAN> and=20
<SPAN class=3Di>mir-16-1</SPAN> map within the intron of a =
non-protein-coding RNA=20
gene of unknown function, which is called <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D8847"><SPAN =

class=3Di>LEU2</SPAN></A>. Clinicians had previously noted that CLL =
patients with=20
13q14 deletions have a more favourable prognosis than those with =
abnormal=20
karyotypes or deletions at other loci such as 11q23 or 17p13. This might =
be due=20
to the fact that homologues of <SPAN class=3Di>mir-15a</SPAN> and <SPAN=20
class=3Di>mir-16-1</SPAN> that are clustered on chromosome 3 (<A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406949"><SPA=
N=20
class=3Di>mir-15b</SPAN></A> and <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406951"><SPA=
N=20
class=3Di>mir-16-2</SPAN></A>, respectively) continue to be expressed at =
low=20
levels in patients with CLL; therefore, 13q14 deletions do not lead to =
the=20
complete elimination of these miRNA families.</P>
<P class=3Dnorm>A recent report by Cimmino <SPAN class=3Di>et al</SPAN>. =
showed that=20
miR-15a and miR-16-1 negatively regulate <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D596"><SPAN=20
class=3Di>BCL2</SPAN></A>, which is an anti-apoptotic gene that is often =

over-expressed in many types of human cancers, including leukaemias and=20
lymphomas<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B94">94=
</A></SUP>.=20
Therefore, it is thought that the deletion or downregulation of <SPAN=20
class=3Di>mir-15a</SPAN> and <SPAN class=3Di>mir-16-1</SPAN> results in =
increased=20
expression of BCL2, promoting leukaemogenesis and lymphomagenesis in=20
haematopoietic cells. The tumour-suppressor role of miR-16 is further =
supported=20
by the finding that two patients with CLL carried a germline C to T =
mutation 7=20
base-pairs downstream of the <SPAN class=3Di>mir-16-1</SPAN> precursor, =
resulting=20
in decreased miR-16-1 expression levels<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B95">95=
</A></SUP>.=20
Although further characterization of the biological functions of miR-15 =
and=20
miR-16-1 have not been completed, evidence indicates that <SPAN=20
class=3Di>mir-16-1</SPAN> expression is preferentially downregulated in =
various=20
leukaemias but not in cancers that originate from other tissues. This =
supports a=20
role for this miRNA in the immune system and B-cell =
differentiation<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B93">93=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B96">96=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B97">97=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B98">98=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B99">99=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B100">1=
00</A></SUP>.</P>
<P class=3Dnorm>Additional studies have shown a strong correlation =
between=20
abrogated expression of miRNAs and oncogenesis. For example, mature =
miRNA levels=20
of miR-143 and <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406937">miR-=
145</A>=20
are significantly reduced in <A=20
href=3D"http://www.cancer.gov/cancertopics/types/colon-and-rectal">colore=
ctal=20
tumours</A><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B97">97=
</A></SUP>.=20
Interestingly, the hairpin precursor transcripts of these miRNAs were =
detected=20
in similar quantities in normal and tumour tissues, indicating that the=20
processing of <SPAN class=3Di>mir-143</SPAN> and <SPAN =
class=3Di>mir-145</SPAN>=20
might be disrupted in these cancer samples. However, the =
tumour-suppressor roles=20
of miR-143 and miR-145 might not be specific to colorectal tissue as =
their=20
expression is also downregulated in breast, prostate, cervical and =
lymphoid=20
cancer cell lines<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B84">84=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B97">97=
</A></SUP>.=20
Another report demonstrated that <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406991">miR-=
21</A>=20
is upregulated in <A=20
href=3D"http://www.cancer.gov/cancertopics/types/brain">glioblastoma</A><=
SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B101">1=
01</A></SUP>.=20
This gene was found to be expressed at a 5=96100-fold higher rate in =
gliomas than=20
in normal tissue. Antisense studies of miR-21 in glioblastoma cell lines =
showed=20
that this miRNA controls cell growth by inhibiting apoptosis but does =
not affect=20
cell proliferation, which implies an oncogenic role for this miRNA. An=20
independent study that used microarray analysis to compare the =
expression of 245=20
miRNAs in glio-blastoma versus normal tissues also identified miR-21 =
levels as=20
being increased in glioblastoma tumours<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B100">1=
00</A></SUP>.=20
However, as <SPAN class=3Di>mir-21</SPAN> is not a brain-specific =
gene<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B100">1=
00</A></SUP>=20
and miR-21 expression is increased in human breast cancer samples<SUP><A =

href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B84">84=
</A></SUP>,=20
this miRNA might have a wider function in tumour progression. More =
studies need=20
to be done to show whether these miRNAs have a direct function in cancer =

progression or are simply differentially modulated in tumours.</P>
<H4 class=3Dnorm id=3DThe-let7-family-negatively-regulates-Ras>The <SPAN =

class=3Di>let-7</SPAN> family negatively regulates Ras</H4>
<P class=3Dnorm>The miRNAs that are encoded by the <SPAN =
class=3Di>let-7</SPAN>=20
family were the first group of oncomirs shown to regulate the expression =
of an=20
oncogene, specifically the Ras genes. Ras proteins are =
membrane-associated=20
GTPase signalling proteins that regulate cellular growth and =
differentiation.=20
About 15=9630% of human tumours possess mutations in Ras genes, and =
activating=20
mutations that result in the increased expression of Ras cause cellular=20
transformation. Therefore, a miRNA that regulates the expression level =
of these=20
potentially oncogenic proteins is predicted to keep the rate of cellular =

proliferation in check. The miRNAs that are encoded by the <SPAN=20
class=3Di>let-7</SPAN> family, which include 12 human homologues, were =
implicated=20
as tumour suppressors because they map to fragile sites associated with =
lung,=20
breast, <A=20
href=3D"http://www.cancer.gov/cancertopics/types/urethral">urothelial</A>=
 and=20
cervical cancers<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B83">83=
</A></SUP>.=20
More direct evidence, presented by Takamizawa <SPAN class=3Di>et =
al</SPAN>.,=20
showed that transcripts of certain <SPAN class=3Di>let-7</SPAN> =
homologues were=20
significantly downregulated in human lung cancer and that this =
correlated with a=20
poor prognosis<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B81">81=
</A></SUP>.=20
These studies further indicate that <SPAN class=3Di>let-7</SPAN> could =
be used=20
diagnostically, as patients with non-small-cell lung carcinomas who =
expressed=20
lower levels of <SPAN class=3Di>let-7</SPAN> had a poorer prognosis and =
shortened=20
post-operative survival. <SPAN class=3Di>In vitro</SPAN> tissue-culture=20
experiments with a human lung adenoma cell line that showed that =
transient=20
delivery of <SPAN class=3Di>let-7</SPAN> can inhibit cellular =
proliferation also=20
indicates that <SPAN class=3Di>let-7</SPAN> probably is a =
tumour-suppressor gene=20
in lung tissue. Therefore, <SPAN class=3Di>let-7</SPAN> could be used=20
therapeutically to treat lung cancer<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B81">81=
</A></SUP>.</P>
<P class=3Dnorm>Clues to how the <SPAN class=3Di>let-7</SPAN> family =
controls=20
cellular proliferation came from studies in <SPAN class=3Di>C. =
elegans</SPAN> in=20
our laboratory. We showed that certain members of the <SPAN =
class=3Di>let-7</SPAN>=20
family genetically interact with oncogenic Ras and that the <SPAN=20
class=3Di>let-7</SPAN> family and Ras are reciprocally expressed. =
Moreover, the 3'=20
UTR of Ras genes contains multiple complementary sites for the <SPAN=20
class=3Di>let-7</SPAN> family<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B60">60=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B82">82=
</A></SUP>.=20
We also found that <SPAN class=3Di>let-7</SPAN> negatively regulates Ras =
in human=20
cells. Overexpression of <SPAN class=3Di>let-7</SPAN> in human cancer =
cell lines=20
results in decreased levels of Ras compared with untreated cells. =
Conversely,=20
the depletion of RNA levels of <SPAN class=3Di>let-7</SPAN> in human =
cancer cell=20
lines that normally express high levels leads to a marked increase in =
Ras=20
protein expression<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B82">82=
</A></SUP>.=20
We also provided evidence that <SPAN class=3Di>let-7</SPAN> directly =
controls Ras=20
expression through 3'-UTR-mediated repression in human cells. Reporter=20
constructs that contain Ras 3' UTRs were downregulated at the =
translational=20
level by <SPAN class=3Di>let-7</SPAN>, and <SPAN class=3Di>let-7</SPAN> =
inhibitors=20
could reverse this repression. Furthermore, when comparing lung tumours =
to=20
normal adjacent cells taken from patients with squamous-cell carcinoma =
of the=20
lung, <SPAN class=3Di>let-7</SPAN> and Ras genes were reciprocally =
expressed:=20
<SPAN class=3Di>let-7</SPAN> homologues were downregulated in the =
tumours, whereas=20
Ras was highly expressed.</P>
<P class=3Dnorm>Taken together, these studies indicate that <SPAN=20
class=3Di>let-7</SPAN> is a promising therapeutic agent to treat lung =
cancers=20
caused by activating mutations in Ras genes. Interestingly, repression =
of <SPAN=20
class=3Di>let-7</SPAN> might be a unique characteristic of lung cancer=20
progression. Analysis of the expression of <SPAN class=3Di>let-7</SPAN> =
in 21=20
different cancer patients, 12 of whom suffered from lung cancer, =
revealed that=20
<SPAN class=3Di>let-7</SPAN> was markedly reduced in all lung cancer =
samples but=20
was only sporadically decreased in other tumour types. In support of =
this=20
finding, microarray analysis of the expression profile of 167 miRNAs in =
lung=20
tumours showed that most miRNA genes were not dramatically modulated in =
lung=20
cancer<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B82">82=
</A></SUP>.</P>
<H4 class=3Dnorm id=3DmiRNAs-associated-with-the-MYC-oncogene>miRNAs =
associated with=20
the <SPAN class=3Di>MYC</SPAN> oncogene</H4>
<P class=3Dnorm>The <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D4609"><SPAN =

class=3Di>MYC</SPAN></A> oncogene, which encodes a basic =
helix-loop-helix=20
transcription factor, is often mutated or amplified in human cancers and =
has=20
been shown to function as an important regulator of cell growth owing to =
its=20
ability to induce both cell proliferation and apoptosis<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B102">1=
02</A></SUP>.=20
Interestingly, there seems to be a close relationship between miRNAs and =
the=20
increased expression of MYC, and this leads to the development of B-cell =

malignancies. For example, an aggressive B-cell leukaemia occurs when =
<SPAN=20
class=3Di>MYC</SPAN> is translocated into the <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406934"><SPA=
N=20
class=3Di>mir-142</SPAN></A> locus. This t(8;17) translocation places =
<SPAN=20
class=3Di>MYC</SPAN> just downstream of the miRNA hairpin sequences and =
under the=20
control of the miRNA promoter<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B41">41=
</A></SUP>.=20
Loss of a conserved region of about 20 nucleotides, which is downstream =
of the=20
<SPAN class=3Di>mir-142</SPAN> precursor, on <SPAN class=3Di>MYC</SPAN>=20
translocation is thought to disrupt miRNA processing, leading to the =
increased=20
expression of MYC and the transformation of B cells.</P>
<P class=3Dnorm>Another miRNA, <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406947">miR-=
155</A>,=20
has also been linked with MYC overexpression and B-cell cancers. This =
miRNA is=20
encoded by nucleotides 241=96262 of <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D114614"><SPA=
N=20
class=3Di>BIC</SPAN></A>. This gene was initially identified as a common =

integration site for the avian leukosis virus and was found to be =
overexpressed=20
in B-cell lymphomas<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B103">1=
03,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B104">1=
04</A></SUP>.=20
For years, researchers were puzzled as to how an RNA that was presumably =
not=20
protein-coding, and was poorly conserved between avian, mouse and human =
genomes,=20
could promote lymphomas in cells that also overexpressed MYC. Metzler =
<SPAN=20
class=3Di>et al</SPAN>. found that the only phylogenetically conserved =
region=20
within <SPAN class=3Di>BIC</SPAN> spans 138 nucleotides and encodes the =
hairpin=20
region of <SPAN class=3Di>mir-155</SPAN> (Ref. <A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B105">1=
05</A>).=20
This group then showed that miR-155 expression is upregulated 100-fold =
in=20
paediatric <A=20
href=3D"http://www.cancer.gov/cancertopics/types/non-hodgkins-lymphoma">B=
urkitt=20
lymphoma</A>, and other studies showed increased miR-155 levels in <A=20
href=3D"http://www.cancer.gov/cancertopics/types/hodgkinslymphoma">Hodgki=
n=20
lymphoma</A> and in primary mediastinal and diffuse large-B-cell =
lymphomas with=20
activated rather than germinal-centre phenotypes<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B106">1=
06,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B107">1=
07,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B108">1=
08</A></SUP>.=20
Therefore, it seems that <SPAN class=3Di>mir-155</SPAN> can function as =
an=20
oncogene in cooperation with <SPAN class=3Di>MYC</SPAN>, whereas it =
normally=20
functions during B-cell selection and possibly targets genes that =
antagonize the=20
MYC pathway.</P>
<P class=3Dnorm>miR-155 has also been reported to be upregulated in =
breast=20
carcinomas, which indicates that there are further roles for this gene =
outside=20
of the haematopoietic system<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B84">84=
</A></SUP>.=20
Furthermore, the organization of the <SPAN class=3Di>mir-155/BIC</SPAN> =
gene is=20
reminiscent of the <SPAN class=3Di>mir-15a</SPAN> and <SPAN=20
class=3Di>mir-16-1</SPAN> genes, which are encoded within the intron of =
the=20
non-protein-coding <SPAN class=3Di>LEU2</SPAN> gene, as well as the <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D407975"><SPA=
N=20
class=3Di>C13orf25</SPAN></A> gene, which contains the <SPAN=20
class=3Di>mir-17=9692</SPAN> cluster that is described below. However, =
the=20
significance of a non-protein-coding RNA harbouring miRNAs within a =
transcript=20
and being linked to oncogenesis is unclear.</P>
<P class=3Dnorm>Two recent papers from He <SPAN class=3Di>et al</SPAN>. =
and=20
O'Donnell <SPAN class=3Di>et al</SPAN>. describe a more direct =
relationship=20
between miRNAs, MYC and cancer<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B109">1=
09,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B110">1=
10</A></SUP>.=20
The 13q31 locus is preferentially amplified in cancers such as diffuse=20
large-B-cell lymphoma, follicular lymphoma, mantle cell lymphoma and =
primary=20
cutaneous B-cell lymphoma. However, the only gene that has been found to =
be=20
upregulated within this amplicon is a non-protein-coding RNA, <SPAN=20
class=3Di>C13orf25</SPAN> (Ref. <A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B11">11=
</A>).=20
This transcript was shown to encode the <SPAN =
class=3Di>mir-17=9692</SPAN> cluster,=20
which includes seven miRNAs: <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406952">miR-=
17-5p</A>,=20
miR-17-3p, <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406953">miR-=
18a</A>,=20
<A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406953">miR-=
19a</A>,=20
<A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406982">miR-=
20a</A>,=20
<A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406980">miR-=
19b-1</A>=20
<A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D407048">miR-=
92-1</A>.=20
He and colleagues analysed cell lines carrying an amplification of 13q31 =
for the=20
expression of 191 miRNAs and found that 6 miRNAs had increased =
expression, which=20
corresponded to more copies of <SPAN class=3Di>C13orf25</SPAN>, and 5 of =
these=20
belonged to the <SPAN class=3Di>mir-17=9692</SPAN> cluster<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B109">1=
09</A></SUP>.=20
This group then showed that the pri-miRNA for the <SPAN =
class=3Di>mir-17=9692</SPAN>=20
cluster was upregulated in 65% of B-cell lymphoma samples that were =
tested. They=20
therefore postulated that increased expression of this cluster =
contributes to=20
cancer formation.</P>
<P class=3Dnorm>To test this directly, the researchers overexpressed the =
<SPAN=20
class=3Di>mir-17=9619b-1</SPAN> cluster (the vertebrate-specific portion =
of the=20
<SPAN class=3Di>mir-17=9692</SPAN> cluster) using a retroviral system in =

haematopoietic stem cells (HSCs) that had been isolated from mice =
carrying the=20
<SPAN class=3Di>Myc</SPAN> transgene. Wild-type mice were given a =
sublethal dose=20
of radiation to wipe out the bone marrow and were then transplanted with =
the=20
retrovirally transduced HSCs. Irradiated animals that received HSCs=20
overexpressing both <SPAN class=3Di>Myc</SPAN> and the <SPAN=20
class=3Di>mir-17=9619b-1</SPAN> cluster developed malignant lymphomas =
faster (<IMG=20
style=3D"BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; VERTICAL-ALIGN: middle; BORDER-RIGHT-WIDTH: =
0px"=20
alt=3Dapprox=20
src=3D"http://www.nature.com/__chars/math/special/sim/black/med/base/glyp=
h.gif">51=20
days) than those animals that received HSCs expressing <SPAN =
class=3Di>Myc</SPAN>=20
alone (3=966 months). Interestingly, similar experiments that used HSCs=20
overexpressing <SPAN class=3Di>Myc</SPAN> in combination with either 96 =
unrelated,=20
single miRNAs or any individual miRNA of the <SPAN =
class=3Di>mir-17=9619b-1</SPAN>=20
cluster did not show accelerated onset of disease. Lymphomas that =
resulted from=20
the increased expression of both MYC and mirRNAs from the <SPAN=20
class=3Di>mir-17=9619b-1</SPAN> cluster had increased cell proliferation =
and lower=20
levels of cell death in comparison with latent malignancies that arose =
from=20
increased MYC expression levels alone, which underwent extensive =
apoptosis.</P>
<P class=3Dnorm>These experiments indicate that miRNAs within the =
truncated <SPAN=20
class=3Di>mir-17=9619b-1</SPAN> cluster function cooperatively as =
oncogenes,=20
possibly by targeting apoptotic factors that are activated in response =
to MYC=20
overexpression. When the brakes of the apoptotic pathway are removed, =
MYC can=20
induce cells to proliferate uncontrollably, which results in cancer. The =

oncogenic function of the <SPAN class=3Di>mir-17=9692</SPAN> cluster is =
further=20
validated by the fact that miR-19a and miR-92-1 expression levels are =
increased=20
in cells from patients with B-cell CLL, that miRNAs from the <SPAN=20
class=3Di>mir-17=9692</SPAN> cluster are overexpressed in lung cancers =
and that=20
<SPAN class=3Di>C13orf25</SPAN> is frequently amplified in alveolar=20
rhabdomyosarcoma and liposarcoma<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B111">1=
11,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B112">1=
12,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B113">1=
13,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B114">1=
14,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B115">1=
15,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B116">1=
16</A></SUP>.</P>
<P class=3Dnorm>O'Donnell <SPAN class=3Di>et al</SPAN>. independently =
identified the=20
<SPAN class=3Di>mir-17=9692</SPAN> cluster as a group of potentially =
cancer-related=20
genes when they carried out a microarray screen of 235 human, mouse and =
rat=20
miRNAs to identify miRNAs that were modulated in a human B-cell line, =
P493-6,=20
that overexpresses MYC<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B110">1=
10</A></SUP>.=20
This study showed that MYC induces the expression of the <SPAN=20
class=3Di>mir-17=9692</SPAN> cluster, and chromatin immunoprecipitation =
experiments=20
revealed that MYC binds within the first intron of <SPAN=20
class=3Di>C13orf25</SPAN>, which implies that MYC directly regulates the =
<SPAN=20
class=3Di>mir-17=9692</SPAN> pri-miRNA transcript. The next step was to =
identify=20
gene targets of the MYC-regulated <SPAN class=3Di>mir-17=9692</SPAN> =
cluster.=20
Previous bioinformatic studies had identified the transcription factor =
<A=20
href=3D"http://us.expasy.org/uniprot/Q01094">E2F1</A> as a target of two =
miRNAs=20
within this cluster =97 miR-17-5p and miR-20a. E2F1 controls the =
transition from=20
G1 to S phase of the cell cycle by regulating genes that are involved in =
DNA=20
replication, cell division and apoptosis<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B117">1=
17</A></SUP>.</P>
<P class=3Dnorm>Interestingly, E2F1 is known to function in a reciprocal =

positive-feedback loop with MYC. O'Donnell <SPAN class=3Di>et al</SPAN>. =
found=20
that inhibition of <SPAN class=3Di>mir-17-5p</SPAN> and <SPAN=20
class=3Di>mir-20a</SPAN> in a cervical cancer cell line markedly =
increased E2F1=20
expression levels without affecting mRNA abundance, which is a hallmark =
of=20
miRNA-mediated gene repression. Mutations of sites in the <SPAN=20
class=3Di>E2F1</SPAN> 3' UTR that are complementary to mir-17-5p and =
mir-20a=20
resulted in increased reporter activity compared with those that result =
from=20
wild-type constructs. Taken together, these studies show that MYC =
induces the=20
transcription of both the <SPAN class=3Di>mir-17=9692</SPAN> cluster and =
<SPAN=20
class=3Di>E2F1</SPAN>, and, in turn, these miRNAs negatively regulate =
the=20
translation of E2F1. Therefore, a model is proposed in which =
MYC-mediated cell=20
growth is tightly regulated by the <SPAN class=3Di>mir-17=9692</SPAN> =
cluster. In=20
the presence of MYC, miRNAs of the <SPAN class=3Di>mir-17=9692</SPAN> =
cluster limit=20
the activity of E2F1 and dampen the cell-proliferative effects of MYC by =

breaking the positive-feedback loop between MYC and E2F1. In this =
context the=20
<SPAN class=3Di>mir-17=9692</SPAN> cluster would function as a tumour =
suppressor,=20
which is in contrast to the findings of He <SPAN class=3Di>et al</SPAN>. =
that are=20
discussed above. Consistent with this model, which was proposed by =
O'Donnell and=20
colleagues, loss of heterozygosity at the 13q31 loci that encodes the =
<SPAN=20
class=3Di>mir-17=9692</SPAN> cluster has been reported in <A=20
href=3D"http://www.cancer.gov/cancertopics/types/liver">hepatocellular=20
carcinomas</A><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B118">1=
18</A></SUP>.=20
However, it has also been reported that although E2F1 drives cell-ular=20
proliferation, when E2F1 expression levels cross a certain threshold, =
excessive=20
levels of this protein induce apoptosis<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B117">1=
17</A></SUP>.=20
In this context, the negative regulation of E2F1 by miRNAs of the <SPAN=20
class=3Di>mir-17=9692</SPAN> cluster might function to block the =
apoptotic activity=20
of E2F1 and stimulate MYC-mediated cellular proliferation, so supporting =
the=20
model that was proposed by He <SPAN class=3Di>et al</SPAN>.</P>
<P class=3Dnorm>The tumour-suppressing and oncogenic nature of the <SPAN =

class=3Di>mir-17=9692</SPAN> cluster emphasizes the complexities of =
cancer=20
progression as well as the intricacies of miRNA-mediated gene =
regulation. These=20
results might also reflect the fact that a single miRNA can control many =

unrelated gene targets, resulting in the control of opposing activities =
such as=20
cellular proliferation and differentiation. Perhaps the ability of the =
<SPAN=20
class=3Di>mir-17=9692</SPAN> cluster to function as either tumour =
suppressors or=20
oncogenes depends on which cell types these miRNAs are expressed in and =
what=20
tissue-specific target mRNAs are present to be regulated. Future studies =
might=20
reveal that miRNAs function as key regulators of many cancer-related =
genes as=20
well as <SPAN class=3Di>BCL2</SPAN>, Ras and <SPAN =
class=3Di>E2F1</SPAN>. In fact, a=20
recent report by Felli <SPAN class=3Di>et al</SPAN>. described the =
ability of <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D407006">miR-=
221</A>=20
and <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D407007">miR-=
222</A>=20
to downregulate the <A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D3815"><SPAN =

class=3Di>KIT</SPAN></A> oncogene to modulate erythropoiesis of =
CD34<SUP>+</SUP>=20
HPCs and inhibit cell growth of an erythroleukaemic TF-1 cell line in a=20
KIT-dependent manner<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B119">1=
19</A></SUP>.=20
In support of the regulatory role of the <SPAN class=3Di>KIT</SPAN> =
oncogene by=20
miRNAs, He <SPAN class=3Di>et al</SPAN>. found that patients with =
papillary=20
thyroid carcinomas showed decreased <SPAN class=3Di>KIT</SPAN> =
transcript and=20
protein levels and reciprocally increased levels of miR-221, miR-222, =
and=20
miR-146 in the tumours, implicating that these miRNAs act as oncogenes =
in this=20
setting<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B132">1=
32</A></SUP>.=20
The study also showed that five out of ten papillary thyroid carcinomas =
showed=20
point mutations in the <SPAN class=3Di>KIT</SPAN> loci specifically =
within the=20
complimentary binding regions for miR-221, miR-222, and miR-146 and =
therefore=20
altered the miRNA=96target interactions in these tumours. More work will =
need to=20
be done in order to understand how these miRNAs contribute to cancer =
progression=20
by downregulating the <SPAN class=3Di>KIT</SPAN> oncogene. Therefore, =
miRNAs might=20
be powerful drug targets that could be used in a broad range of cancer=20
therapies.</P>
<H4 class=3Dnorm id=3DmiRNA-profiling-might-aid-cancer-diagnosis>miRNA =
profiling=20
might aid cancer diagnosis</H4>
<P class=3Dnorm>Northern-blot analyses and miRNA microarrays have been =
useful in=20
determining tissue-specific 'signatures' of miRNA genes in humans<SUP><A =

href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B37">37=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B44">44=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B46">46=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B96">96=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B120">1=
20,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B121">1=
21,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B122">1=
22,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B123">1=
23,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B124">1=
24,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B125">1=
25,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B126">1=
26,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B127">1=
27,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B128">1=
28</A></SUP>.=20
The distinct miRNA-expression profiles that have been observed in =
certain organs=20
underscore the importance of miRNAs in stem-cell maintenance and in =
directing=20
the differentiation of certain cell types during development. =
Researchers are=20
now using miRNA-expression signatures to classify cancers and to define =
miRNA=20
markers that might predict favourable prognosis<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B81">81=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B84">84=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B95">95=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B98">98=
,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B100">1=
00,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B101">1=
01,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B109">1=
09,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B110">1=
10,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B112">1=
12</A></SUP>.</P>
<P class=3Dnorm>A recent report from Lu <SPAN class=3Di>et al</SPAN>. =
found that the=20
expression profiles of relatively few miRNAs (<IMG=20
style=3D"BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; VERTICAL-ALIGN: middle; BORDER-RIGHT-WIDTH: =
0px"=20
alt=3Dapprox=20
src=3D"http://www.nature.com/__chars/math/special/sim/black/med/base/glyp=
h.gif">200=20
genes), were required to accurately classify human cancers<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B98">98=
</A></SUP>.=20
This group designed a novel method, which is based on <A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/glossary/nrc1840.html#df2=
">bead-based=20
flow cytometry</A>, to study the expression of miRNAs in both normal and =

cancerous tissues. Tumours that originated from various tissues were =
clustered=20
solely on the basis of their miRNA-expression profiles. This resulted in =
tumours=20
being grouped according to their embryonic lineage. For example, tumours =
of=20
endothelial origin such as colon, liver, <A=20
href=3D"http://www.cancer.gov/cancertopics/types/pancreatic">pancreas</A>=
 and <A=20
href=3D"http://www.cancer.gov/cancertopics/types/stomach">stomach</A> =
were=20
clustered together, and tumours of haematopoietic origin also clustered=20
together. Interestingly, when the expression profiles of about 16,000=20
protein-coding mRNA genes were analysed in the same tumour dataset, =
hierarchical=20
clustering was unable to group tumours in a coherent manner =97 for =
example,=20
gastrointestinal-derived tumours could not be grouped together. =
Therefore, miRNA=20
signatures within tumours reflect their developmental history, a =
hypothesis that=20
is compatible with the evidence that miRNAs direct tissue-specific =
developmental=20
functions.</P>
<P class=3Dnorm>These researchers also compared the miRNA-expression =
profiles of=20
tumour samples versus normal tissue and found that 129 out of 217 miRNAs =
had a=20
lower level of expression in tumours regardless of the tissue of origin. =

Therefore, in a global sense, miRNAs might function to drive cells into =
a more=20
differentiated state, and the expression profiles of miRNAs in tumours =
compared=20
with normal tissues might represent the degree of differentiation in =
those=20
cells. These studies establish miRNAs as 'oncomirs' and imply that =
abnormalities=20
in miRNA expression might directly result in the de-differentiation of =
cells,=20
allowing tumour formation to occur.</P>
<P class=3Dnorm>An established library of miRNA signatures or =
miRNA-expression=20
profiles, assigned for each class of tumour, might help both the =
diagnosis and=20
treatment of cancer. These libraries could easily be established as =
miRNAs can=20
be isolated from formalin-fixed paraffin-embedded archival =
tissues<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B120">1=
20</A></SUP>.=20
The differential expression of certain miRNAs has already been shown to =
be an=20
accurate predictor of a patient's overall prognosis. From a therapeutic=20
standpoint, miRNA-expression profiling might prove to be a powerful tool =
for=20
clinicians when defining a treatment regimen for their patients. As =
proof of=20
principle, Lu <SPAN class=3Di>et al</SPAN>. found that miRNA signatures =
could=20
successfully categorize poorly differentiated tumours that were =
considered=20
histologically non-diagnostic into their appropriate tissue-specific=20
lineages<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B98">98=
</A></SUP>.</P>
<H4 class=3Dnorm id=3DFuture-approaches-using-miRNA-therapies>Future =
approaches=20
using miRNA therapies</H4>
<P class=3Dnorm>The emergence of miRNAs as important cancer-prevention =
genes is=20
likely to have a large effect on gene therapies that are designed to =
block=20
tumour progression. Large-scale expression screens, similar to those =
described=20
above that compare miRNA levels in tumours versus normal tissues, will =
be useful=20
in identifying novel miRNAs that are involved in cancer. Also, =
functional=20
screens that are designed to select miRNA genes that specifically =
control=20
cancer-related processes such as cell proliferation and apoptosis, =
similar to=20
those that were conducted by Cheng <SPAN class=3Di>et al</SPAN>.<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B99">99=
</A></SUP>,=20
will also help in this search. In the future, the administration of =
synthetic=20
anti-sense oligonucleotides that encode sequences that are complementary =
to=20
mature oncogenic miRNAs =97 termed <A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/glossary/nrc1840.html#df3=
">anti-miRNA=20
oligonucleotides</A> (AMOs) =97 might effectively inactivate miRNAs in =
tumours and=20
slow their growth. In the clinic, miRNA inactivation could be =
accomplished=20
through frequent or continuous delivery of 2'-<SPAN =
class=3Di>O</SPAN>-methyl or=20
locked-nucleic-acid antisense oligonucleotides, which are specifically =
designed=20
to be more stable and less toxic than other cancer treatments in order =
to target=20
transforming miRNAs such as <SPAN class=3Di>mir-155</SPAN>.</P>
<P class=3Dnorm>The use of antagomirs, which are AMOs that are =
conjugated with=20
cholesterol, has recently emerged as an effective approach to inhibit =
miRNA=20
activity in various organs when injected into mice, and might be =
promising=20
therapeutic agents<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B129">1=
29</A></SUP>.=20
Conversely, techniques to overexpress miRNAs that function as tumour=20
suppressors, such as those that are encoded by the <SPAN =
class=3Di>let-7</SPAN>=20
family, could be used to treat specific tumour types. Transient =
expression=20
systems that use viral or liposomal delivery might be useful approaches =
for=20
administering large quantities of miRNAs. These techniques would involve =

expressing the pre-miRNA hairpin and flanking sequences, potentially =
under the=20
control of tissue-specific promoters, to stimulate the endogenous miRNA=20
machinery to direct proper miRNA processing and gene-targeted =
repression.=20
However, employing similar methods that use siRNAs as cancer gene =
therapies have=20
shown that the immune response can limit the effectiveness of RNA=20
delivery<SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B130">1=
30,=20
</A></SUP><SUP><A=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#B131">1=
31</A></SUP>.=20
More development of these methods is needed before miRNA treatments can =
move=20
from the laboratory bench to the bedside. Whether miRNAs become the =
'magic=20
bullet' of the future remains to be seen, but research in this area will =

undoubtedly provide insight into the underlying mechanisms of=20
oncogenesis.</P></DIV>
<DIV id=3Dlinks>
<H4>Links</H4>
<H5>DATABASES</H5>
<H6><A href=3D"http://www.cancer.gov/">National Cancer =
Institute</A></H6>
<UL>
  <LI class=3Dfirst><A=20
  href=3D"http://www.cancer.gov/cancertopics/types/breast">breast =
cancer</A>=20
  <LI><A=20
  =
href=3D"http://www.cancer.gov/cancertopics/types/non-hodgkins-lymphoma">B=
urkitt=20
  lymphoma</A>=20
  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/cervical">cervical=20
  cancer</A>=20
  <LI><A=20
  =
href=3D"http://www.cancer.gov/cancertopics/types/colon-and-rectal">colore=
ctal=20
  tumours</A>=20
  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/brain">glioblastoma</A> =

  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/liver">hepatocellular=20
  carcinoma</A>=20
  <LI><A=20
  =
href=3D"http://www.cancer.gov/cancertopics/types/hodgkinslymphoma">Hodgki=
n=20
  lymphoma</A>=20
  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/leukemia">leukaemia</A> =

  <LI><A href=3D"http://www.cancer.gov/cancertopics/types/lung">lung =
cancer</A>=20
  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/myeloma">multiple=20
  myeloma</A>=20
  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/ovarian">ovarian=20
  cancer</A>=20
  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/pancreatic">pancreatic=20
  cancer</A>=20
  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/prostate">prostate=20
  cancer</A>=20
  <LI><A href=3D"http://www.cancer.gov/cancertopics/types/kidney">renal=20
  tumours</A>=20
  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/stomach">stomach=20
  cancer</A>=20
  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/testicular">testicular=20
  cancer</A>=20
  <LI><A =
href=3D"http://www.cancer.gov/cancertopics/types/urethral">urothelial=20
  cancer</A>=20
  <LI><A href=3D"http://www.cancer.gov/cancertopics/types/wilms">Wilms =
tumour</A>=20
  </LI></UL>
<H6><A=20
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene">Entrez-G=
ene</A></H6>
<UL>
  <LI class=3Dfirst><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D3772493"><SP=
AN=20
  class=3Di>let-7</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D192669"><SPA=
N=20
  class=3Di>AGO3</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D26523"><SPAN=
=20
  class=3Di>AGO1</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D192670"><SPA=
N=20
  class=3Di>AGO4</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D7490">WT1</A=
>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D9271"><SPAN =

  class=3Di>HIWI</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D34521"><SPAN=
=20
  class=3Di>piwi</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D3772549"><SP=
AN=20
  class=3Di>bantam</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D3772293">miR=
-14</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D3565055">miR=
-273</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D178087"><SPA=
N=20
  class=3Di>lys-6</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D387176">miR-=
181</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D494324">miR-=
375</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406935">miR-=
143</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406911"><SPA=
N=20
  class=3Di>mir-125b-1</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406948"><SPA=
N=20
  class=3Di>mir-15a</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406950"><SPA=
N=20
  class=3Di>mir-16-1</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D8847"><SPAN =

  class=3Di>LEU2</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406949"><SPA=
N=20
  class=3Di>mir-15b</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406951"><SPA=
N=20
  class=3Di>mir-16-2</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D596"><SPAN=20
  class=3Di>BCL2</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406937">miR-=
145</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406991">miR-=
21</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D4609"><SPAN =

  class=3Di>MYC</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406934"><SPA=
N=20
  class=3Di>mir-142</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406947">miR-=
155</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D114614"><SPA=
N=20
  class=3Di>BIC</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D407975"><SPA=
N=20
  class=3Di>C13orf25</SPAN></A>=20
  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406952">miR-=
17-5p</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406953">miR-=
18a</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406953">miR-=
19a</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406982">miR-=
20a</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D406980">miR-=
19b-1</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D407048">miR-=
92-1</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D407006">miR-=
221</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D407007">miR-=
222</A>=20

  <LI><A=20
  =
href=3D"http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=3Dgene&amp;amp;c=
md=3DRetrieve&amp;amp;dopt=3Dfull_report&amp;amp;list_uids=3D3815"><SPAN =

  class=3Di>KIT</SPAN></A> </LI></UL>
<H6><A href=3D"http://us.expasy.org/uniprot/">UniProtKB</A></H6>
<UL>
  <LI class=3Dfirst><A =
href=3D"http://us.expasy.org/uniprot/Q9NRR4">Drosha</A>=20
  <LI><A href=3D"http://us.expasy.org/uniprot/Q8WYQ5">Pasha</A>=20
  <LI><A href=3D"http://us.expasy.org/uniprot/Q9ULC9">exportin 5</A>=20
  <LI><A href=3D"http://us.expasy.org/uniprot/Q9UPY3">Dicer</A>=20
  <LI><A href=3D"http://us.expasy.org/uniprot/Q01094">E2F1</A> =
</LI></UL>
<H5>FURTHER INFORMATION</H5>
<UL>
  <LI class=3Dfirst><A=20
  href=3D"http://www.mcdb.yale.edu/facultystaff/slack.html">Frank =
Slack's=20
  homepage</A>=20
  <LI><A =
href=3D"http://www.ncbi.nlm.nih.gov/projects/genome/guide/human">Human=20
  Genome Resource</A>=20
  <LI><A href=3D"http://microrna.sanger.ac.uk/">miRBase</A>=20
  <LI><A href=3D"http://genes.mit.edu/mirscan">miRScan</A>=20
  <LI><A href=3D"http://www.microrna.org/">miRanda</A>=20
  <LI><A href=3D"http://genes.mit.edu/targetscan">TargetScan</A> =
</LI></UL></DIV>
<DIV id=3Dacknowledgements><A class=3Dbacktotop=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#top">To=
p<SPAN=20
class=3Dhidden> of page</SPAN></A>
<H3>Acknowledgements</H3>
<P class=3Dnorm>We thank Helge Grosshans and Diya Banerjee for critical =
reading of=20
the manuscript. F.J.S. is supported by grants from the National Science=20
Foundation and the National Institutes of Health and A.E.-K. is =
supported by a=20
Ruth L. Kirschstein National Research Service Award Individual =
Postdoctoral=20
Fellowship.</P>
<H4 class=3Dinline>Competing interests statement </H4>
<P class=3Dfollows-h4>The authors declare no competing financial=20
interests.</P></DIV>
<DIV id=3DReferences><A class=3Dbacktotop=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#top">To=
p<SPAN=20
class=3Dhidden> of page</SPAN></A>
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<DIV id=3Daffiliations-notes>
<H3>Author affiliations</H3>
<OL>
  <LI id=3Da1>Yale University, Department of Molecular, Cellular &amp;=20
  Developmental Biology, 266 Whitney Avenue, New Haven, Connecticut =
06520,=20
  USA.</LI></OL>
<P class=3Dcaff>Correspondence to: Frank J. Slack<SUP><A =
title=3D"affiliated with "=20
href=3D"http://www.nature.com/nrc/journal/v6/n4/full/nrc1840.html#a1">1</=
A></SUP>=20
Email:&nbsp;<A=20
href=3D"mailto:frank.slack@yale.edu">frank.slack@yale.edu</A></P></DIV>
<DIV class=3Dno-header><A class=3Dbacktotop=20
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Content-Type: text/css;
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Content-Transfer-Encoding: quoted-printable
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------=_NextPart_000_0000_01C8B1D0.A3459760
Content-Type: text/css;
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------=_NextPart_000_0000_01C8B1D0.A3459760
Content-Type: text/css;
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Content-Transfer-Encoding: quoted-printable
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PADDING-BOTTOM: 4px; MARGIN: 15px 0px 0px
}
SPAN.booktitle {
	FONT-STYLE: italic
}
SPAN.journalname {
	FONT-STYLE: italic
}
SPAN.journalnumber {
	FONT-WEIGHT: bold
}
SPAN.tm {
	VERTICAL-ALIGN: super
}
LEGEND {
	COLOR: #000
}
* HTML #breadcrumb DIV {
	MARGIN-TOP: 0px; PADDING-LEFT: 5px; PADDING-TOP: 1px
}
#breadcrumb DIV {
	CLEAR: left; MARGIN-TOP: 0px; PADDING-LEFT: 5px; BACKGROUND: #fff; =
PADDING-BOTTOM: 10px; PADDING-TOP: 1px
}
#breadcrumb DIV A:link {
	FONT-SIZE: 70%; COLOR: #000; TEXT-DECORATION: none
}
#breadcrumb DIV A:visited {
	FONT-SIZE: 70%; COLOR: #000; TEXT-DECORATION: none
}
#breadcrumb DIV A:hover {
	COLOR: #000; TEXT-DECORATION: underline
}
#breadcrumb SPAN.divider {
	FONT-SIZE: 70%; COLOR: #000
}
#breadcrumb SPAN.thisitem {
	FONT-SIZE: 70%; COLOR: #000
}
#login-inpage {
	MARGIN: 0px; COLOR: #000; BACKGROUND-COLOR: #eaeaea
}
#login-inpage A {
	FONT-SIZE: 70%
}
#login-inpage DIV.userpassbox {
	FLOAT: left; WIDTH: 8em
}
#login-inpage .passdetails {
	CLEAR: left
}
#login-inpage INPUT.text {
	FONT-SIZE: 80%; MARGIN: 0px; WIDTH: 9em; COLOR: #000
}
#login-inpage LABEL {
	FONT-SIZE: 70%
}
#login-inpage INPUT.button {
	MARGIN-TOP: 5px
}
#login-inpage #savepass-inpage {
	MARGIN-BOTTOM: -2px
}
#login-inpage DIV.error {
	PADDING-LEFT: 20px; FONT-SIZE: 80%; BACKGROUND: =
url(/common/images/login_error.gif) no-repeat 0px 4px; MARGIN-BOTTOM: =
5px; COLOR: #c30
}
#login-inpage DIV.error SPAN {
	FONT-WEIGHT: bold
}
#head-er HR {
	DISPLAY: none
}
#head-er .header-nonad-wrapper {
	PADDING-BOTTOM: 15px; BORDER-BOTTOM: #fff 1px solid
}
#head-er .accessibility-login-wrap {
	FLOAT: left; WIDTH: 100%; BORDER-BOTTOM: #000 1px solid; =
BACKGROUND-COLOR: #eaeaea
}
#head-er .accessibility-login {
	PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 5px; =
PADDING-TOP: 4px; TEXT-ALIGN: right
}
#head-er .accessibility-login A.nature-link {
	FLOAT: left; FONT: bold 160%/80% Garamond, serif; WIDTH: 4.9em; COLOR: =
#b1062e; MARGIN-RIGHT: 20px; TEXT-ALIGN: left; TEXT-DECORATION: none
}
* HTML #head-er .accessibility-login A.nature-link:hover {
	BACKGROUND: none transparent scroll repeat 0% 0%
}
#head-er .accessibility-login A.nature-link:hover {
	BACKGROUND: =
url(/common/includes/header/images/nature.com.underline.gif) no-repeat =
left bottom
}
#head-er .accessibility-login IMG.nature-logo {
	BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; FLOAT: left; =
BORDER-BOTTOM-WIDTH: 0px; MARGIN: 1px 20px 0px 0px; WIDTH: 82px! =
important; HEIGHT: 11px; BORDER-RIGHT-WIDTH: 0px
}
#head-er .accessibility-login .jumplinks {
	PADDING-RIGHT: 0px; PADDING-LEFT: 0px; FLOAT: left; PADDING-BOTTOM: =
0px; MARGIN: 0px; WIDTH: 45%; PADDING-TOP: 0px; LIST-STYLE-TYPE: none; =
TEXT-ALIGN: left
}
#head-er .accessibility-login .jumplinks LI {
	DISPLAY: inline; PADDING-LEFT: 15px
}
#head-er .accessibility-login .jumplinks LI.first {
	PADDING-LEFT: 0px
}
#head-er .accessibility-login .jumplinks A {
	PADDING-LEFT: 10px; BACKGROUND: =
url(/common/images/icons/b1062e/arrow_down.gif) no-repeat left center; =
COLOR: #000; TEXT-DECORATION: none
}
#head-er .accessibility-login .jumplinks A:hover {
	TEXT-DECORATION: underline
}
#head-er DIV.logon {
=09
}
#head-er .logon .onlyformacie {
	FONT-SIZE: 3px
}
#head-er .logon .onlyformacie {
	DISPLAY: none
}
#head-er .logon A.login {
	PADDING-RIGHT: 15px; PADDING-LEFT: 0px; FONT-WEIGHT: bold; BACKGROUND: =
url(/common/includes/header/images/logon.gif) no-repeat right center; =
PADDING-BOTTOM: 0px; TEXT-TRANSFORM: uppercase; COLOR: #b1062e; =
PADDING-TOP: 0px; LETTER-SPACING: -0.1em; TEXT-DECORATION: none
}
#head-er .logon A.login:hover {
	BACKGROUND: url(/common/includes/header/images/logon_over.gif) =
no-repeat right center; TEXT-DECORATION: underline
}
#head-er .logon A.login:active {
	BACKGROUND: url(/common/includes/header/images/logon.gif) no-repeat =
right center
}
#head-er .logon A.logoff {
	PADDING-RIGHT: 15px; PADDING-LEFT: 15px; FONT-WEIGHT: bold; BACKGROUND: =
url(/common/includes/header/images/logoff.gif) no-repeat right center; =
PADDING-BOTTOM: 0px; TEXT-TRANSFORM: uppercase; COLOR: #b1062e; =
PADDING-TOP: 0px; LETTER-SPACING: -0.1em; TEXT-DECORATION: none
}
#head-er .logon P {
	MARGIN: 0px
}
#head-er .logon .user {
	FONT-WEIGHT: bold
}
#head-er .logon .inst {
	FONT-WEIGHT: bold
}
#head-er .image-user-services-wrap {
	FLOAT: left; WIDTH: 100%; BACKGROUND-COLOR: #d6335c
}
#head-er .image-user-services {
	BORDER-BOTTOM: transparent 1px solid
}
* HTML #head-er .image-user-services {
	BORDER-BOTTOM-WIDTH: 0px
}
#head-er .image-user-services IMG.journal-header {
	BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; FLOAT: left; =
BORDER-BOTTOM-WIDTH: 0px; MARGIN: 0px; WIDTH: 344px; HEIGHT: 74px; =
BORDER-RIGHT-WIDTH: 0px
}
#head-er .user-services {
	FLOAT: right
}
#head-er .div1.user-services {
	WIDTH: 12.35em! important
}
#head-er .div2.user-services {
	WIDTH: 8.7em! important
}
#head-er .user-services UL {
	BORDER-RIGHT: #fff 0px solid; PADDING-RIGHT: 0px; BORDER-TOP: #fff 1px =
solid; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: 22px 0px 21px; =
BORDER-LEFT: #fff 1px solid; PADDING-TOP: 0px; BORDER-BOTTOM: #fff 0px =
solid; LIST-STYLE-TYPE: none; TEXT-ALIGN: right
}
#head-er .user-services UL.list1 {
	BORDER-RIGHT: #fff 1px solid
}
#head-er .user-services LI {
	BORDER-BOTTOM: #fff 1px solid
}
#head-er .user-services A {
	PADDING-RIGHT: 13px; DISPLAY: block; PADDING-LEFT: 0px; BACKGROUND: =
url(/common/images/icons/fff/arrow_s.gif) #767474 no-repeat right =
center; PADDING-BOTTOM: 1px; TEXT-TRANSFORM: uppercase; COLOR: #fff; =
PADDING-TOP: 1px; TEXT-DECORATION: none
}
#head-er .user-services A:hover {
	BACKGROUND: url(/common/images/icons/fff/arrow_s.gif) #000 no-repeat =
right center
}
#head-er .publications-search-wrap {
	BACKGROUND: #000
}
* HTML #head-er .publications-search {
	PADDING-BOTTOM: 3px; PADDING-TOP: 3px
}
#head-er .publications-search {
	PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 4px; =
PADDING-TOP: 4px
}
* HTML #head-er .publications {
	MARGIN: 6px 0px 0px
}
#head-er .publications {
	PADDING-RIGHT: 0px; PADDING-LEFT: 0px; FLOAT: left; PADDING-BOTTOM: =
0px; MARGIN: 4px 0px 0px; PADDING-TOP: 0px; LIST-STYLE-TYPE: none
}
#head-er .publications LI {
	DISPLAY: inline; MARGIN-LEFT: 10px
}
#head-er .publications LI.first {
	MARGIN-LEFT: 0px
}
#head-er .publications A {
	PADDING-RIGHT: 15px; BACKGROUND: =
url(/common/images/icons/fff/arrow_s.gif) no-repeat right center; =
TEXT-TRANSFORM: uppercase; COLOR: #fff; TEXT-DECORATION: none
}
#head-er .publications A:hover {
	TEXT-DECORATION: underline
}
#head-er .search {
	TEXT-ALIGN: right
}
#head-er .search FORM {
	DISPLAY: inline; MARGIN: 0px
}
#head-er .search FORM A {
	PADDING-RIGHT: 14px; BACKGROUND: =
url(/common/images/icons/fff/arrow_s.gif) no-repeat right center; =
TEXT-TRANSFORM: uppercase; COLOR: #fff; TEXT-DECORATION: none
}
#head-er .search FORM A:hover {
	TEXT-DECORATION: underline
}
#head-er #searchtext {
	WIDTH: 10em
}
#head-er .publications-search LABEL {
	TEXT-TRANSFORM: uppercase; COLOR: #fff
}
#head-er INPUT {
	FONT-SIZE: 100%
}
#head-er SELECT {
	FONT-SIZE: 100%
}
#head-er .publications-search INPUT.button {
	PADDING-RIGHT: 3px; PADDING-LEFT: 3px; FONT-WEIGHT: bold; BACKGROUND: =
#eaeaea; PADDING-BOTTOM: 0px; COLOR: #000; PADDING-TOP: 0px
}
#head-er .accessibility-login {
	PADDING-RIGHT: 1%; PADDING-LEFT: 1%
}
#head-er .image-user-services {
	PADDING-RIGHT: 1%; PADDING-LEFT: 1%
}
#head-er .publications-search {
	PADDING-RIGHT: 1%; PADDING-LEFT: 1%
}
.header-leaderboard-wrap {
	PADDING-RIGHT: 1%; PADDING-LEFT: 1%; BACKGROUND: =
url(/common/includes/header/images/bg_leaderboard.gif) no-repeat 50% =
0px; OVERFLOW: hidden
}
.header-leaderboard {
	MAX-HEIGHT: 105px; TEXT-ALIGN: center
}
* HTML .header-leaderboard IMG {
	BORDER-BOTTOM-WIDTH: 1px
}
* HTML .header-leaderboard ilayer {
	BORDER-BOTTOM-WIDTH: 1px
}
* HTML .header-leaderboard IFRAME {
	BORDER-BOTTOM-WIDTH: 1px
}
* HTML .header-leaderboard EMBED {
	BORDER-BOTTOM-WIDTH: 1px
}
.header-leaderboard IMG {
	BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; =
BORDER-LEFT-WIDTH: 0px; MIN-HEIGHT: 90px; MAX-WIDTH: 728px; =
PADDING-BOTTOM: 0px; PADDING-TOP: 0px; BORDER-BOTTOM: #fff 1px solid; =
HEIGHT: 90px; BORDER-RIGHT-WIDTH: 0px
}
.header-leaderboard ilayer {
	BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; =
BORDER-LEFT-WIDTH: 0px; MIN-HEIGHT: 90px; MAX-WIDTH: 728px; =
PADDING-BOTTOM: 0px; PADDING-TOP: 0px; BORDER-BOTTOM: #fff 1px solid; =
HEIGHT: 90px; BORDER-RIGHT-WIDTH: 0px
}
.header-leaderboard IFRAME {
	BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; =
BORDER-LEFT-WIDTH: 0px; MIN-HEIGHT: 90px; MAX-WIDTH: 728px; =
PADDING-BOTTOM: 0px; PADDING-TOP: 0px; BORDER-BOTTOM: #fff 1px solid; =
HEIGHT: 90px; BORDER-RIGHT-WIDTH: 0px
}
.header-leaderboard EMBED {
	BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; =
BORDER-LEFT-WIDTH: 0px; MIN-HEIGHT: 90px; MAX-WIDTH: 728px; =
PADDING-BOTTOM: 0px; PADDING-TOP: 0px; BORDER-BOTTOM: #fff 1px solid; =
HEIGHT: 90px; BORDER-RIGHT-WIDTH: 0px
}
.header-leaderboard TABLE {
	BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; =
BORDER-LEFT-WIDTH: 0px; MAX-WIDTH: 728px; PADDING-BOTTOM: 0px; =
MARGIN-LEFT: auto; WIDTH: 100%; MARGIN-RIGHT: auto; PADDING-TOP: 0px; =
BORDER-BOTTOM: #fff 1px solid; BORDER-RIGHT-WIDTH: 0px
}
* HTML .header-leaderboard TABLE IMG {
	HEIGHT: auto
}
#constrain {
	MARGIN-TOP: 14px
}
#global-subject-areas {
	BORDER-RIGHT: #ebebeb 1px solid; BORDER-TOP: #ebebeb 1px solid; =
BACKGROUND: #ebebeb; MARGIN: 0px 0px 20px; BORDER-LEFT: #ebebeb 1px =
solid; BORDER-BOTTOM: #ebebeb 1px solid
}
#global-subject-areas ABBR {
	BORDER-TOP-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: =
0px; BORDER-RIGHT-WIDTH: 0px; TEXT-DECORATION: none
}
#global-subject-areas A:link {
	FONT-WEIGHT: bold; COLOR: #9d0006; TEXT-DECORATION: none
}
#global-subject-areas A:visited {
	FONT-WEIGHT: bold; COLOR: #9d0006; TEXT-DECORATION: none
}
#global-subject-areas A:hover {
	TEXT-DECORATION: underline
}
#global-subject-areas H2 {
	PADDING-RIGHT: 10px; PADDING-LEFT: 10px; FONT-SIZE: 110%; BACKGROUND: =
#b1062e; PADDING-BOTTOM: 5px; MARGIN: 0px; COLOR: #fff; PADDING-TOP: =
5px; BORDER-BOTTOM: #fff 1px solid
}
#global-subject-areas H3 {
	PADDING-RIGHT: 10px; PADDING-LEFT: 10px; FONT-SIZE: 110%; BACKGROUND: =
#b1062e; PADDING-BOTTOM: 5px; MARGIN: 0px; COLOR: #fff; PADDING-TOP: =
5px; BORDER-BOTTOM: #fff 1px solid
}
#global-subject-areas H4 {
	PADDING-RIGHT: 10px; PADDING-LEFT: 10px; FONT-SIZE: 110%; BACKGROUND: =
#b1062e; PADDING-BOTTOM: 5px; MARGIN: 0px; COLOR: #fff; PADDING-TOP: =
5px; BORDER-BOTTOM: #fff 1px solid
}
#global-subject-areas H5 {
	PADDING-RIGHT: 10px; PADDING-LEFT: 10px; FONT-SIZE: 110%; BACKGROUND: =
#b1062e; PADDING-BOTTOM: 5px; MARGIN: 0px; COLOR: #fff; PADDING-TOP: =
5px; BORDER-BOTTOM: #fff 1px solid
}
#global-subject-areas H3.view {
	PADDING-RIGHT: 17px; PADDING-LEFT: 17px; FONT-SIZE: 100%; BACKGROUND: =
url(/common/images/bg.7x1.b1062e.gif) #fff repeat-y left top; =
PADDING-BOTTOM: 5px; MARGIN: 0px; COLOR: #9d0006; PADDING-TOP: 5px; =
BORDER-BOTTOM: #fff 1px solid
}
#global-subject-areas H4.view {
	PADDING-RIGHT: 17px; PADDING-LEFT: 17px; FONT-SIZE: 100%; BACKGROUND: =
url(/common/images/bg.7x1.b1062e.gif) #fff repeat-y left top; =
PADDING-BOTTOM: 5px; MARGIN: 0px; COLOR: #9d0006; PADDING-TOP: 5px; =
BORDER-BOTTOM: #fff 1px solid
}
#global-subject-areas H5.view {
	PADDING-RIGHT: 17px; PADDING-LEFT: 17px; FONT-SIZE: 100%; BACKGROUND: =
url(/common/images/bg.7x1.b1062e.gif) #fff repeat-y left top; =
PADDING-BOTTOM: 5px; MARGIN: 0px; COLOR: #9d0006; PADDING-TOP: 5px; =
BORDER-BOTTOM: #fff 1px solid
}
#global-subject-areas UL {
	PADDING-RIGHT: 0px; PADDING-LEFT: 0px; FONT-SIZE: 100%; PADDING-BOTTOM: =
0px; MARGIN: 0px; PADDING-TOP: 0px; LIST-STYLE-TYPE: none
}
#global-subject-areas UL UL {
	FONT-SIZE: 100%
}
#global-subject-areas UL LI {
	PADDING-RIGHT: 5px; PADDING-LEFT: 5px; PADDING-BOTTOM: 5px; =
LINE-HEIGHT: 150%; PADDING-TOP: 5px; BORDER-BOTTOM: #fff 1px solid
}
#global-subject-areas UL LI LI {
	PADDING-RIGHT: 0px; PADDING-LEFT: 5px; BORDER-BOTTOM-WIDTH: 0px; =
PADDING-BOTTOM: 0px; PADDING-TOP: 0px
}
#global-subject-areas UL LI H4 {
	BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; =
FONT-WEIGHT: normal; BORDER-LEFT-WIDTH: 0px; FONT-SIZE: 100%; =
BACKGROUND: none transparent scroll repeat 0% 0%; BORDER-BOTTOM-WIDTH: =
0px; PADDING-BOTTOM: 0px; MARGIN: 0px; COLOR: #666; PADDING-TOP: 0px; =
FONT-STYLE: italic; BORDER-RIGHT-WIDTH: 0px
}
#global-subject-areas UL LI H5 {
	BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; =
FONT-WEIGHT: normal; BORDER-LEFT-WIDTH: 0px; FONT-SIZE: 100%; =
BACKGROUND: none transparent scroll repeat 0% 0%; BORDER-BOTTOM-WIDTH: =
0px; PADDING-BOTTOM: 0px; MARGIN: 0px; COLOR: #666; PADDING-TOP: 0px; =
FONT-STYLE: italic; BORDER-RIGHT-WIDTH: 0px
}
#global-subject-areas UL LI H6 {
	BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; =
FONT-WEIGHT: normal; BORDER-LEFT-WIDTH: 0px; FONT-SIZE: 100%; =
BACKGROUND: none transparent scroll repeat 0% 0%; BORDER-BOTTOM-WIDTH: =
0px; PADDING-BOTTOM: 0px; MARGIN: 0px; COLOR: #666; PADDING-TOP: 0px; =
FONT-STYLE: italic; BORDER-RIGHT-WIDTH: 0px
}
MAP.journal-nav {
	DISPLAY: block; MARGIN-BOTTOM: 20px
}
MAP.journal-nav UL {
	PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: =
0px; PADDING-TOP: 0px; LIST-STYLE-TYPE: none; TEXT-ALIGN: left
}
MAP.journal-nav UL LI {
	MARGIN: 0px; BORDER-BOTTOM: #fff 1px solid; BACKGROUND-COLOR: #eaeaea
}
MAP.journal-nav UL LI.grouped {
	BORDER-BOTTOM: #eaeaea 1px solid
}
* HTML MAP.journal-nav UL LI A {
	PADDING-RIGHT: 5px; PADDING-LEFT: 5px; PADDING-BOTTOM: 3px; =
PADDING-TOP: 2px
}
* HTML MAP.journal-nav UL LI A:visited {
	PADDING-RIGHT: 5px; PADDING-LEFT: 5px; PADDING-BOTTOM: 3px; =
PADDING-TOP: 2px
}
MAP.journal-nav UL LI A {
	PADDING-RIGHT: 5px; DISPLAY: block; PADDING-LEFT: 5px; FONT-WEIGHT: =
bold; FONT-SIZE: 80%; PADDING-BOTTOM: 4px; MARGIN: 0px; COLOR: #000; =
PADDING-TOP: 4px; TEXT-DECORATION: none
}
MAP.journal-nav UL LI A:visited {
	PADDING-RIGHT: 5px; DISPLAY: block; PADDING-LEFT: 5px; FONT-WEIGHT: =
bold; FONT-SIZE: 80%; PADDING-BOTTOM: 4px; MARGIN: 0px; COLOR: #000; =
PADDING-TOP: 4px; TEXT-DECORATION: none
}
MAP.journal-nav UL LI A:hover {
	COLOR: #fff; BACKGROUND-COLOR: #c00; TEXT-DECORATION: none
}
MAP.journal-nav UL LI A.on {
	COLOR: #fff; BACKGROUND-COLOR: #c00
}
MAP.journal-nav UL LI A.on:visited {
	COLOR: #fff; BACKGROUND-COLOR: #c00
}
MAP.journal-nav UL LI A.submission {
	PADDING-LEFT: 20px! important; BACKGROUND: =
url(/common/images/online_submissions.gif) no-repeat left center
}
MAP.journal-nav UL LI UL.subnav {
	BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; =
PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-TOP: 0px; LIST-STYLE-TYPE: =
none; TEXT-ALIGN: left
}
MAP.journal-nav UL LI UL.subnav LI {
	BACKGROUND: url(/common/images/nav_dots2.gif) #fafafa no-repeat 11px =
5px; BORDER-BOTTOM-WIDTH: 0px; MARGIN: 0px
}
MAP.journal-nav UL LI UL.subnav LI.on {
	BACKGROUND: url(/common/images/nav_dots1.gif) #c00 no-repeat 11px 5px; =
BORDER-BOTTOM: #fafafa 1px solid
}
* HTML MAP.journal-nav UL LI UL.subnav A {
	PADDING-RIGHT: 10px! important; PADDING-LEFT: 20px! important; =
PADDING-BOTTOM: 2px! important; PADDING-TOP: 2px! important
}
MAP.journal-nav UL LI UL.subnav A:link {
	PADDING-RIGHT: 10px; DISPLAY: block; PADDING-LEFT: 20px; FONT-WEIGHT: =
normal; FONT-SIZE: 70%; PADDING-BOTTOM: 2px; MARGIN: 0px; COLOR: #000; =
PADDING-TOP: 2px; TEXT-DECORATION: none
}
MAP.journal-nav UL LI UL.subnav A:visited {
	PADDING-RIGHT: 10px; DISPLAY: block; PADDING-LEFT: 20px; FONT-WEIGHT: =
normal; FONT-SIZE: 70%; PADDING-BOTTOM: 2px; MARGIN: 0px; COLOR: #000; =
PADDING-TOP: 2px; TEXT-DECORATION: none
}
MAP.journal-nav UL LI UL.subnav A:hover {
	BACKGROUND: url(/common/images/nav_dots1.gif) #fab934 no-repeat 11px =
5px; COLOR: #fff; TEXT-DECORATION: none
}
MAP.journal-nav UL LI UL.subnav A.on {
	COLOR: #fff
}
MAP.journal-nav UL LI UL.subnav A.on:visited {
	COLOR: #fff
}
#journalnav MAP.supplementary UL A {
	FONT-WEIGHT: normal; FONT-SIZE: 70%
}
#journalnav MAP.supplementary UL A:visited {
	FONT-WEIGHT: normal; FONT-SIZE: 70%
}
H2.society-resources {
	PADDING-RIGHT: 5px; PADDING-LEFT: 5px; FONT-WEIGHT: bold; FONT-SIZE: =
80%; PADDING-BOTTOM: 2px; MARGIN: 0px; COLOR: #fff; PADDING-TOP: 2px; =
BACKGROUND-COLOR: #fab934
}
UL.society-resources {
	PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: 0px =
0px 20px; PADDING-TOP: 0px; LIST-STYLE-TYPE: none; TEXT-ALIGN: left
}
UL.society-resources LI {
	MARGIN: 0px; BACKGROUND-COLOR: #eaeaea
}
UL.society-resources A {
	PADDING-RIGHT: 5px! important; DISPLAY: block; PADDING-LEFT: 5px! =
important; FONT-SIZE: 70%; PADDING-BOTTOM: 3px! important; MARGIN: 0px; =
COLOR: #000; PADDING-TOP: 3px! important; TEXT-DECORATION: none
}
UL.society-resources A:visited {
	PADDING-RIGHT: 5px! important; DISPLAY: block; PADDING-LEFT: 5px! =
important; FONT-SIZE: 70%; PADDING-BOTTOM: 3px! important; MARGIN: 0px; =
COLOR: #000; PADDING-TOP: 3px! important; TEXT-DECORATION: none
}
UL.society-resources A:hover {
	COLOR: #fff; BACKGROUND-COLOR: #fab934; TEXT-DECORATION: none
}
DIV.society-logo {
	MARGIN-BOTTOM: 20px; TEXT-ALIGN: center
}
DIV.society-logo IMG {
	WIDTH: 100%
}
H2.npg-resources {
	PADDING-RIGHT: 5px; PADDING-LEFT: 5px; FONT-WEIGHT: bold; FONT-SIZE: =
80%; PADDING-BOTTOM: 2px; MARGIN: 0px; COLOR: #fff; PADDING-TOP: 2px; =
BACKGROUND-COLOR: #b1062e
}
UL.npg-resources {
	PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: 0px =
0px 20px; PADDING-TOP: 0px; LIST-STYLE-TYPE: none; TEXT-ALIGN: left
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#foot-er #footer-copyright P {
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------=_NextPart_000_0000_01C8B1D0.A3459760
Content-Type: text/css;
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Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.nature.com/reviews/style/group.css

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DIV#breadcrumb DIV {
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------=_NextPart_000_0000_01C8B1D0.A3459760
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.nature.com/common/scripts/global.js

(function($) {
	$(document).ready(function () {

		// enables dropdown ads to display whilst maintaining .user-services =
visibility aat small resolutions
		// yes, it involves browser detection, and yes, that is truly =
horrible.
		if (jQuery.browser.mozilla) {
	=09
			var dropdownAd =3D false;
	=09
			// for each descendant of .leaderboard that isn't a <script>, =
determine if it's taller than standard leaderboard height
			$("#hdr .leaderboard *:not(script):not(.padder)").each(function() {
				if($(this).height() > 90) {
					dropdownAd =3D true;
				}
			});
	=09
			if (dropdownAd) {
				$("#hdr .leaderboard").css("overflow","visible");
				$("#hdr .leaderboard .padder").css("overflow","visible");
				$("#hdr .leaderboard").hover(function() {
					$("#hdr .leaderboard").css("overflow","visible");
					$("#hdr .leaderboard .padder").css("overflow","visible");
				},function() {
					$("#hdr .leaderboard").css("overflow","hidden");
					$("#hdr .leaderboard .padder").css("overflow","hidden");
				});		=09
			}
		}
	=09
	});
}) (jQuery);
------=_NextPart_000_0000_01C8B1D0.A3459760--

