From: Subject: BioMed Central | Abstract | Berberine enhances inhibition of glioma tumor cell migration and invasiveness mediated by arsenic trioxide Date: Fri, 18 Apr 2008 19:24:03 +0200 MIME-Version: 1.0 Content-Type: multipart/related; type="text/html"; boundary="----=_NextPart_000_0043_01C8A189.C32FCB60" X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3198 This is a multi-part message in MIME format. ------=_NextPart_000_0043_01C8A189.C32FCB60 Content-Type: text/html; charset="utf-8" Content-Transfer-Encoding: quoted-printable Content-Location: http://www.biomedcentral.com/1471-2407/8/58/abstract/ =EF=BB=BF BioMed Central | = Abstract | Berberine enhances inhibition of glioma tumor cell migration = and invasiveness mediated by arsenic trioxide

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Research article

Berberine enhances inhibition of glioma tumor cell migration = and=20 invasiveness mediated by arsenic trioxide

Tseng-Hsi Lin^*=20¹^,3 ,=20 Hsing-Chun Kuo^{* =}¹^,4 ,=20 Fen-Pi Chou¹ and = Fung-Jou Lu²

¹Institute of Biochemistry and Biotechnology, Chung = Shan=20 Medical University, Taichung 402, Taiwan

²School of Applied Chemistry, Chung Shan Medical = University,=20 Taichung, Taiwan

³Division of Hematology, Department of Internal = Medicine,=20 Taichung Veterans General Hospital, Taichung, Taiwan

⁴Department of Medical Research, Chang Gung Memorial = Hospital, Kaohsiung Medical Center, Kaohsiung 833, = Taiwan

=20 author email=20 corresponding author email* = Contributed=20 equally

BMC Cancer 2008, = 8:58doi:10.1186/1471-2407-8-58


Published:	25 February 2008

<= A=20 name=3Dabstract>

Abstract

Background

Arsenic trioxide (As₂O₃) exhibits = promising=20 anticarcinogenic activity in acute promyelocytic leukemic patients = and=20 induces apoptosis in various tumor cells in vitro. Here, = we=20 investigated the effect of the natural alkaloid berberine on=20 As₂O₃-mediated inhibition of cancer cell = migration=20 using rat and human glioma cell lines.

Methods

The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium = bromide (MTT)=20 assay was used to determine the viability of rat C6 and human U-87 = glioma=20 cells after treatment with As₂O₃or = berberine, and=20 after co-treatment with As₂O₃and berberine. = The=20 wound scratch and Boyden chamber assays were applied to determine = the=20 effect of As₂O₃and berberine on the = migration=20 capacity and invasiveness of glioma cancer cells. Zymography and = Western=20 blot analyses provided information on the effect of = As₂O₃₌₂₀and berberine on the intracellular translocation and = activation of=20 protein kinase C (PKC), and some PKC-related downstream factors. = Most=20 assays were performed three times, independently, and data were = analyzed=20 using ANOVA.

Results

The cell viability studies demonstrated that berberine enhances = As₂O₃-mediated inhibition of glioma cell = growth=20 after 24 h incubation. Untreated control cells formed a confluent = layer,=20 the formation of which was inhibited upon incubation with 5 =CE=BCM As₂O₃. The = latter effect was=20 even more pronounced in the presence of 10 =CE=BCM=20 berberine. The As₂O₃-mediated reduction in = motility=20 and invasion of glioma cells was enhanced upon co-treatment with=20 berberine. Furthermore, it has been reported that PKC isoforms = influence=20 the morphology of the actin cytoskeleton, as well as the = activation of=20 metalloproteases MT1-MMP and MMP-2, reported to be involved in = cancer cell=20 migration. Treatment of glioma cells with As₂O_{3 =}and=20 berberine significantly decreased the activation of PKC =CE=B1 and =CE=B5 and led to actin = cytoskeleton rearrangements.=20 The levels of two downstream transcription factors, myc and jun, = and=20 MT1-MMP and MMP-2 were also significantly reduced.

Conclusion

Upon co-treatment of glioma cells with As₂O_{3 =}and=20 berberine, cancer cell metastasis can be significantly inhibited, = most=20 likely by blocking the PKC-mediated signaling pathway involved in = cancer=20 cell migration. This study is potentially interesting for the = development=20 of novel chemotherapeutic approaches in the treatment of malignant = gliomas=20 and cancer development in = general.

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=C2=A9 1999-2008 BioMed Central Ltd = unless otherwise=20 stated =

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window.open(url,n,'width=3D'+wi+',height=3D'+he+',top=3D10,left=3D10,resi= zable=3Dyes,scrollbars=3Dyes,toolbar=3Dyes,location=3Dyes,directories=3Dy= es,status=3Dyes,menubar=3Dyes,copyhistory=3Dyes'); } function loadhalf(width) { var popwidth =3D typeof width !=3D 'undefined'? width+40 : 640; if (popwidth > screen.availWidth) popwidth =3D screen.availWidth; if (popwidth < 640) popwidth =3D 640; window.focus(); window.moveTo(0,0); window.resizeTo(popwidth,screen.availHeight); window.blur; } function loadfull()=20 { window.focus(); window.moveTo(0,0); window.resizeTo(screen.availWidth,screen.availHeight); window.blur; } function loadInParent(link)=20 { if (parent.window.opener.closed) window.open(link); else { parent.window.opener.location =3D link; parent.window.focus(); } } function toggleauthors(auth,button) { try{ var aut=3D document.getElementById(auth); var but =3D document.getElementById(button); if (aut.style.display !=3D "none") { aut.style.display =3D "none"; but.src =3D "/bmcimages/expand2.gif"; } else { aut.style.display =3D "block"; but.src =3D "/bmcimages/contract2.gif"; } } catch(e){} } function andtotext() { document.write(" | Return to text") } function stepback(text) { document.write("" + text + "") } function turn() { if (window.location.hash.length > 0) window.history.back();=20 else window.location=3D"#"; } function goback() { window.history.back();=20 } // Movie embedding. Part 1 - Player detection (incl. version detection), = courtesy of http://skyzyx.com/ // QuickTime detection var quicktime=3Dnew Object(); // Set some base values quicktime.installed=3Dfalse; quicktime.version=3D'0.0'; // for non-IE browsers if (navigator.plugins && navigator.plugins.length) { for (x=3D0; x=3D 0.0; movietext=3D"

"; break; case "mp4": cond =3D document.getElementById && quicktime.version >=3D 6.0; movietext=3D"

"; break; case "peg": ; case "mpg": cond =3D document.getElementById && quicktime.version >=3D 6.4; movietext=3D"

"; break; case "avi": cond =3D document.getElementById && quicktime.version >=3D 3.0; movietext=3D"

"; break; case "mov": cond =3D document.getElementById && quicktime.version >=3D 3.0 ; movietext=3D"

"; break; } =09 // This executes the replacement if (cond) { document.getElementById(id).innerHTML=3Dmovietext; link =3D false; } } //Functions related to FLV movies // Stopping the movie when another is started var currentItem; function getUpdate(typ,pr1,pr2,pid) { if (typ =3D=3D "state") { =09 // Play=20 if (pr1 =3D=3D 2) { =09 // Pause active movie if it is playing. if (currentItem !=3D null && currentItem !=3D pid) { thisMovie(currentItem).sendEvent('playpause'); } =09 // Save current movie currentItem =3D pid;=09 =09 // Flush active movie var on Stop (3) and Pause (0). } else if(pr1 =3D=3D 3 || pr1 =3D=3D 0) { = =20 currentItem =3D null; }=20 } }; // This is a javascript handler for the player and is always needed. function thisMovie(movieName) { if(navigator.appName.indexOf("Microsoft") !=3D -1) { return window[movieName]; } else { return document[movieName]; } }; // Replacing the jpeg with the movie var link =3D true; function playvideo(id,moviename,width,height) { if (document.getElementById) { var flvheight =3D height + 20; var so =3D new = SWFObject('/movies/mediaplayer.swf','js'+id,width,flvheight,'7'); =20 so.useExpressInstall('/movies/expressinstall.swf');=20 so.addParam('allowfullscreen','true'); so.addParam('salign','lt'); so.addParam('quality','high'); so.addParam('scale','noscale'); so.addVariable('file',moviename); =20 so.addVariable('autostart','true'); so.addVariable('showdigits','false'); so.addVariable('useaudio','false'); so.addVariable('usefullscreen','true'); so.addVariable('showicons','false'); so.addVariable('enablejs','true'); so.addVariable('javascriptid','js'+id); = =20 so.addVariable('width',width); so.addVariable('height',flvheight); so.write(id); link =3D false; } } /** * SWFObject v1.5: Flash Player detection and embed - = http://blog.deconcept.com/swfobject/ * * SWFObject is (c) 2007 Geoff Stearns and is released under the MIT = License: * http://www.opensource.org/licenses/mit-license.php * */ if(typeof deconcept=3D=3D"undefined"){var deconcept=3Dnew = Object();}if(typeof = deconcept.util=3D=3D"undefined"){deconcept.util=3Dnew = Object();}if(typeof = deconcept.SWFObjectUtil=3D=3D"undefined"){deconcept.SWFObjectUtil=3Dnew = Object();}deconcept.SWFObject=3Dfunction(_1,id,w,h,_5,c,_7,_8,_9,_a){if(!= document.getElementById){return;}this.DETECT_KEY=3D_a?_a:"detectflash";th= is.skipDetect=3Ddeconcept.util.getRequestParameter(this.DETECT_KEY);this.= params=3Dnew Object();this.variables=3Dnew = Object();this.attributes=3Dnew = Array();if(_1){this.setAttribute("swf",_1);}if(id){this.setAttribute("id"= ,id);}if(w){this.setAttribute("width",w);}if(h){this.setAttribute("height= ",h);}if(_5){this.setAttribute("version",new = deconcept.PlayerVersion(_5.toString().split(".")));}this.installedVer=3Dd= econcept.SWFObjectUtil.getPlayerVersion();if(!window.opera&&document.all&= &this.installedVer.major>7){deconcept.SWFObject.doPrepUnload=3Dtrue;}if(c= ){this.addParam("bgcolor",c);}var = q=3D_7?_7:"high";this.addParam("quality",q);this.setAttribute("useExpress= Install",false);this.setAttribute("doExpressInstall",false);var = _c=3D(_8)?_8:window.location;this.setAttribute("xiRedirectUrl",_c);this.s= etAttribute("redirectUrl","");if(_9){this.setAttribute("redirectUrl",_9);= }};deconcept.SWFObject.prototype=3D{useExpressInstall:function(_d){this.x= iSWFPath=3D!_d?"expressinstall.swf":_d;this.setAttribute("useExpressInsta= ll",true);},setAttribute:function(_e,_f){this.attributes[_e]=3D_f;},getAt= tribute:function(_10){return = this.attributes[_10];},addParam:function(_11,_12){this.params[_11]=3D_12;= },getParams:function(){return = this.params;},addVariable:function(_13,_14){this.variables[_13]=3D_14;},g= etVariable:function(_15){return = this.variables[_15];},getVariables:function(){return = this.variables;},getVariablePairs:function(){var _16=3Dnew Array();var = key;var _18=3Dthis.getVariables();for(key in = _18){_16[_16.length]=3Dkey+"=3D"+_18[key];}return = _16;},getSWFHTML:function(){var = _19=3D"";if(navigator.plugins&&navigator.mimeTypes&&navigator.mimeTypes.l= ength){if(this.getAttribute("doExpressInstall")){this.addVariable("MMplay= erType","PlugIn");this.setAttribute("swf",this.xiSWFPath);}_19=3D"

0){_19+=3D"flashvar= s=3D\""+_1c+"\"";}_19+=3D"/>";}else{if(this.getAttribute("doExpressInstal= l")){this.addVariable("MMplayerType","ActiveX");this.setAttribute("swf",t= his.xiSWFPath);}_19=3D"

";}return = _19;},write:function(_20){if(this.getAttribute("useExpressInstall")){var = _21=3Dnew = deconcept.PlayerVersion([6,0,65]);if(this.installedVer.versionIsValid(_21= )&&!this.installedVer.versionIsValid(this.getAttribute("version"))){this.= setAttribute("doExpressInstall",true);this.addVariable("MMredirectURL",es= cape(this.getAttribute("xiRedirectUrl")));document.title=3Ddocument.title= .slice(0,47)+" - Flash Player = Installation";this.addVariable("MMdoctitle",document.title);}}if(this.ski= pDetect||this.getAttribute("doExpressInstall")||this.installedVer.version= IsValid(this.getAttribute("version"))){var n=3D(typeof = _20=3D=3D"string")?document.getElementById(_20):_20;n.innerHTML=3Dthis.ge= tSWFHTML();return = true;}else{if(this.getAttribute("redirectUrl")!=3D""){document.location.r= eplace(this.getAttribute("redirectUrl"));}}return = false;}};deconcept.SWFObjectUtil.getPlayerVersion=3Dfunction(){var = _23=3Dnew = deconcept.PlayerVersion([0,0,0]);if(navigator.plugins&&navigator.mimeType= s.length){var x=3Dnavigator.plugins["Shockwave = Flash"];if(x&&x.description){_23=3Dnew = deconcept.PlayerVersion(x.description.replace(/([a-zA-Z]|\s)+/,"").replac= e(/(\s+r|\s+b[0-9]+)/,".").split("."));}}else{if(navigator.userAgent&&nav= igator.userAgent.indexOf("Windows CE")>=3D0){var axo=3D1;var = _26=3D3;while(axo){try{_26++;axo=3Dnew = ActiveXObject("ShockwaveFlash.ShockwaveFlash."+_26);_23=3Dnew = deconcept.PlayerVersion([_26,0,0]);}catch(e){axo=3Dnull;}}}else{try{var = axo=3Dnew = ActiveXObject("ShockwaveFlash.ShockwaveFlash.7");}catch(e){try{var = axo=3Dnew ActiveXObject("ShockwaveFlash.ShockwaveFlash.6");_23=3Dnew = deconcept.PlayerVersion([6,0,21]);axo.AllowScriptAccess=3D"always";}catch= (e){if(_23.major=3D=3D6){return _23;}}try{axo=3Dnew = ActiveXObject("ShockwaveFlash.ShockwaveFlash");}catch(e){}}if(axo!=3Dnull= ){_23=3Dnew deconcept.PlayerVersion(axo.GetVariable("$version").split(" = ")[1].split(","));}}}return = _23;};deconcept.PlayerVersion=3Dfunction(_29){this.major=3D_29[0]!=3Dnull= ?parseInt(_29[0]):0;this.minor=3D_29[1]!=3Dnull?parseInt(_29[1]):0;this.r= ev=3D_29[2]!=3Dnull?parseInt(_29[2]):0;};deconcept.PlayerVersion.prototyp= e.versionIsValid=3Dfunction(fv){if(this.majorfv.major){return = true;}if(this.minorfv.minor){return true;}if(this.rev=3D0;i--){_2f[i].style.display=3D"none";for(var x in = _2f[i]){if(typeof = _2f[i][x]=3D=3D"function"){_2f[i][x]=3Dfunction(){};}}}};if(deconcept.SWF= Object.doPrepUnload){if(!deconcept.unloadSet){deconcept.SWFObjectUtil.pre= pUnload=3Dfunction(){__flash_unloadHandler=3Dfunction(){};__flash_savedUn= loadHandler=3Dfunction(){};window.attachEvent("onunload",deconcept.SWFObj= ectUtil.cleanupSWFs);};window.attachEvent("onbeforeunload",deconcept.SWFO= bjectUtil.prepUnload);deconcept.unloadSet=3Dtrue;}}if(!document.getElemen= tById&&document.all){document.getElementById=3Dfunction(id){return = document.all[id];};}var = getQueryParamValue=3Ddeconcept.util.getRequestParameter;var = FlashObject=3Ddeconcept.SWFObject;var SWFObject=3Ddeconcept.SWFObject; 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