gpc3过表达对肝癌huh7细胞生物学功能影响的研究

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1、Researchontheeffectofbiologicalfunctionbyup-regulating【objectiveJexpressionofGPC3inHuh．7cellsAbstractGPC3isakindofheparansulfatemembraneglycoprotein，andParticularlyhighexpressioninlivercancertissue．Atpresent，mostscholarsdiscoverthatsignificantlychangesuppressedoftumorbiologybehaviorbyinhibiting

2、GPC3expressioninlivercells，thusthinkGPC3isakindofpromotecancergenesinlivercancercells．ButGPC3wasfoundthatcancauseproteinexpressionmissingduetothemethylationinpromoterinmesothelioma，ovariancancercellline，breastcancerandlungcalqcer,implyingGPC3mayplayaroleoftumorsuppressorgenes，andalsothesimilarp

3、henomenonwasfoundinstudyoflivercancerThisresearchchoicehuh-7cellwhichishighexpressionofGPC3gene，anddetectthechangesofhuh一7cellbiologyfunctionafterup—regulatingtheexpressionofGPC3，inordertoresearchhowGPC3playsaroleoftumorsuppressorgenesinlivercancerIMethods】1．TransfectingGPC3geneexpressionofeuka

4、ryoticexpressionplasmidintoHuh-7cellsbyLiposomemethod2．DetectingtheexpressionofGPC3mRNAandproteinbyusingfluorescencequantitativePCRandWesternBlot，validatingofGPC3geneexpression．3．DetectingcellcycleandcellapoptosisbyusingtheAnnexinV-F1TC／PIdoublestainflowcytometry4．Detectingcellproliferationbyus

5、ingEDU5．DetectingcellmigrationandinvasionabilitybyusingTranswellmethodfResultsJ1．ConstructingGPC3genewasclonedintotheeukaryoticexpressionvectorpcDNA3．1+GPC3sequencing，F：5’GATTCAGCCTTGGACATCAATG3’，whichissameasthatoftheoriginalsequence，indicatingsuccessfulconstructionofGPC3eukaryoticexpressionve

6、ctorpcDNA3．1+，whichcanbeusedforsubsequentexpressionexperiments．2．GPC3expressionvectorpcDNA3．1+transienttransfectionHuh一7cellsafter72h，fluorescencequantitativePCRandWesternBlotdetectiondisplayedGPC3mRNAandproteinexpressionweresignificantlyhigherthanthecontrolgroup，indicatingsuccessfulincrementof

7、Huh一7cellsofGPC3expressionlevel，enablingtocarryoutsubsequentresearchandexperimentofcellbiologicalbehavior．3．InoverexpressionofHuh一7cellsgroup，Huh一7cellgroupandNCgroup，thenumberofcellsinG1phasewere(49．77士2．08)％，(49．28士1．0)％and(50．2