论著基因芯片技术分析斑蝥素对肝癌细胞细胞毒作用的分子机

13,2,3,2,3,3,1(1.,200438;2.,200433;3.,200433)[]::12.5LmolöLQGY770324h:(p27ref21DNApolymerasedeltaXRCC9)(malatedehydrogenaseADPöATPtranslocase)(c2myctre)(bladdercancerrelatedprotein),(BCRA2BTG2dual2specificityproteinphosphatase)(ATL2derivedPMA2responsivepeptide):[];;;;QGY7703[]R735.7[]A[]02582879X(2003)0620645205Genemicroarraysindetectingmolecularmechanismsofcantharidin-mediatedcytotoxicityonhumanhepaticcancercellsHUHe2Ping13,ZHANGJun2Ping2,YINGKang3,XIAOZhen2Yu2,WUHong2Mei3,MAOYu2Min3,WUMeng2Chao1(1.DepartmentofGastroenterology,EasternHepatobiliarySurgeryHospital,SecondMilitaryMedicalUniversity,Shanghai200438,China;2.DepartmentofPharmacology,SchoolofPharmacy,SecondMilitaryMedicalUniversity,Shanghai200433;3.StateKeyLaboratoryofGeneticEngineering,SchoolofLifeScience,FudanUniversity,Shanghai200433)[ABSTRACT]Objective:Tostudythemolecularmechanismsofantitumoragentcantharidinonhumancancercells.Meth2ods:GeneexpressionprofilewasidentifiedbycDNAmicroarraysinhepaticcancerQGY27703cellsexposedtocantharidin(12.5LmolöL)for24h.Results:Cantharidin2treatedcellshaddecreasedexpressionofgenescodingforproteinsinvolvedincellcycleprogress(p27,DNApolymerasedelta,XRCC,ref21),energymetabolism(malatedehydrogenase,ADPöATPtranslo2case),oncogenicactivation(c2myc,tre)andtumor2specificexpression(bladdercancerrelatedprotein).Incontrast,thesetreatedcellsoverexpressedseveralgenesencodingintracellulargrowth2inhibitoryproteins(BCRA2,BTG2,dual2specificityproteinphosphatase)andproapoptoticgenes(ATL2derivedPMA2responsivepeptide).Conclusion:Thesefindingssuggestthatalter2ationsinspecificgenesinvolvedinmodulatingthecellcycleprogress,cellproliferation,energymetabolism,andapoptosiscas2cadesmayberesponsibleforcantharidin2mediatedcytotoxicity.[KEYWORDS]cantharidin;hepaticcancer;cells;geneexpressionprofiling;cDNAmicroarray;QGY7703cells[AcadJSecMilMedUniv,2003,24(6):6452649](cantharidin,CTD),,,[1,2],[3],[4,5]QGY7703,11.1QGY7703,10%(GibcoBRL)RPMI1640(Sigma),75ml,90%,,10%(0.1%Me2SO),CTD(Sigma,12.5LmolöL),37,5%CO224h1.2RNARNA,OligotexmRNAkit(Qiagen)mRNA,RiboGreenRNAquantitationkit(Molecular546AcadJSecMilMedUniv2003Jun;24(6)[](ZX01B05).[](19612),(),,,.3Correspondingauthor.E2mail:[e-mail]©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.Probes,Eugene,OR)mRNAmRNAcDNAdUTP(Cy52dUTPöCy32dUTP)cDNA,12800cDNA(UnitedGeneHold2ings,Ltd.,PRC)4216hBioDoorScanArray3000(GeneralScanning),ImaGene3.0(BioDiscovery,Inc.)Cy3Cy5[7]2-2-222.1,QGY7703,DNARNA(1)1Tab1EffectsofcantharidinonexpressionofgenesinvolvedincellcycleprogressGeneAccessionnumberFunctionFoldchangeMitosisChromatin2specifictranscriptionelongationfactorNM2007192Transcriptionfactor-3.9P27AB001740Kinetochore-3.3Generaltranscriptionfactor3A(GTF3A)D32257Transcriptionfactor-2.5Chromatinassemblyfactor2ÉP150U20979Chromatinassembly-2.4KinesinsuperfamilymotorKIF4AF071592Kinetochore-2.2CentromereautoantigenCM95724Kinetochore-2.0DNAreplicationDinb1AB027564DNApolymerase-5.5DNAligaseÉM36067DNAligase-3.7DNApolymerasedeltaU21090DNApolymeraseandexonuclease-3.5DihydropteridinereductaseM16447Nucleotidesynthesis-3.4DNApolymeraseepsilonl09561DNApolymerase-3.2Primase,polypeptide1(49000)NM2000946DNAprimase-3.0RibonucleotidereductaseM1X59543Nucleotidesynthesis-2.9MethylenetetrahydrofolatedehydrogenaseJ04031Folatemetabolismregulator-2.5PutativegenecontainingtheMCM3AL034343DNAreplicationlicensingfactor-2.5ReplicationproteinA(70000)M63488Single2strandedDNAbindingprotein-2.4CTPsynthetaseX52142Nucleotidesynthesis-2.3ReplicationproteinA(32000)J05249Single2strandedDNAbindingprotein-2.1ReplicationfactorC(36000)L07540PCNAclampformation-2.0DNArepairRef21(APEXnuclease)S43127Apurinicendonuclease-7.5Xrcc9NM2004629DNApostreplicationrepairorcell-6.0cyclecheckpointcontrolErcc2X52221CorrectingthesensitivitytoUVradiationand-5.5defectivenucleotideexcisionrepairReca2likeprotein(Hrec2)U92074DNArecombinationandrepair-3.7DNA2dependentproteinkinaseU47077Double2strandbreakrepair-3.5catalyticsubunit(DNA2Pkcs)andV(D)JrecombinationMuts,E.coli,homologof,6(Hmsh6)U73737Mismatch2bindingfactor-3.4RNAprocessingandproteinsynthesisnucleolinM60858Rrnatranscriptionalcontrol,ribosome-6.9maturationandassembly,andribosomalcomponentstransportation40SribosomalproteinS3X55715RibosomalproteinandDNArepair-5.0Histidyl2trnasynthetasehomolog(HO3)U18937Proteinsynthesis-4.2Cdna(weaklysimilartoputativepre2mrnaAK001751RNAsplicing-3.8splicingfactorRNAhelicase)Pre2mrnasplicingfactorSF2p32M69039RNAsplicing-3.5Putativetrnasynthetase2likeproteinU07424Proteinsynthesis-3.464620036,24©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.()GeneAccessionnumberFunctionFoldchangeSmallnuclearribonucleoproteinpolypeptideA(SNRPA1)NM2003090Proteinsynthesis-3.2RNApolymeraseÊU37689RNAtranscription-2.9Valyl2Trnasynthetase2(VARS2)NM2006295Proteinsynthesis-2.8RibosomalproteinS14M13934Proteinsynthesis-2.7Nucleolarproteinhnop56Y12065Ribosomebiogenesis-2.5RibosomalproteinL3X06323Proteinsynthesis-2.3Eukaryotictranslationinitiationfactor(Eif3)U78525Proteinsynthesis-2.3Smallnuclearrib