《P21-activated kinase 5 plays essential roles in the proliferation and tumorigenicity of human hepatocellular carcinoma.》.pdf

ActaPharmacologicaSinica (2014)35：82—88 @2014CPSandSIMM Allrightsreserved1671—4083／14 $32．O0 WWW．nature．com／aps OriginalArticle P21-activatedkinase5playsessentialrolesin theproliferationandtumorigenicityofhuman hepat0CellUIarcarcinoma Zhe—pingFANG ，，Bei—geJIANG ，，Xue—fengGU。·，BinZHAO。，Rui．1iangGE，，Fa—biaoZHANG ， DepartmentofHepatobiliarySurgery,TaizhouHospitalofZhejiangProvince，WenzhouMedicalcollege，Linhai317000，China； 。EasternHepatobiliarySurgeryHospital，SecondMilitaryMedicalUniversity,Shanghai200438，China；。ClinicalResearchCenter, AffiliatedHospitalofGuangdongMedicalCollege,Zhanjiang524001，China Aim：T0investigatetherolesofP21一activatedkinase5(PAK5)inproliferationandtumorigenicityofhumanhepatocellularcarcinoma (HCC)． Methods：HCCandmatchedparaneoplastictistissuesampleswereobtainedfr0m 3Opatients．HumanHCCcelllinesSMMC7721． HepG2，Hep3B，SK—HEP一1，Huh一7，andlivercell¨neHL-7702wereexamined．TheexpressionofPAK5genewasstudiedusingreal-time qPCRandWesternblotting．CelIproliferationwasquantifiedwiththeMTI-assay．CelIcyclewasanalyzedwithflowcytometry．The tumorigenicityofLv—shRNA-transfectedHepG2cellswasevaluatedinBALB／cAnudemice． Results：ThemRNAIeveIofPAK5wassignificantlyhigherin25outof30HCCsamplescomparedtothematchedDarane0pIastic tissues．TheHCCcelIlinesshowedvaryingexpressionofPAK5protein．andthehighestIeve1wasfoundintheHepG2cells．PAK5gene silencinginHepG2cellsmarkedlyreducedthecellproliferationandcolonyformation．andinducedcellcyclearrestintheG phase． Furthermore，PAK5geResilencingsuppressedthetumorformationinnudemice，andsignificantlydecreasedtheexpressionofHCC． relatedgenesCyclinD1andbeta—catenin． Conclusion：PAK5mayplayessentialrolesintheinitiationandprogressionofhumanHCC．Thus．itmaybeaneffectivetherapeutic targetorperhapsserveasaclinicaldiagnosticorprognosticmarkerinhumanHCC． Keywords：P21-activatedkinase5fPAK5)；humanhepatocellularcarcinoma；HepG2cell；CyclinD1；beta—catenin；cellc