《过表达转录因子21抑制SMMC-7721肝癌细胞的增殖和迁移并促进其凋亡》.pdf

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1、884细胞与分子免疫学杂志(ChinJCellMolImmuno1)2015，31(7·论著·文章编号：1007—8738(2015)07—0884—05过表达转录因子21抑制SMMC-7721肝癌细胞的增殖和迁移并促进其凋亡谭婧宇，张国庆，刘锐，周密，李中堂，吴忠均(重庆医科大学附属第一医院肝胆外科，重庆400016)[摘要]目的探讨转录因子21(TCF21)基因对SMMC-7721肝癌细胞增殖、迁移和凋亡的影响及机制。方法采用脂质体将pcDNA3．1(+)TCF21及pcDNA3．1(+)质

2、粒稳定转染入SMMC-7721细胞，分别为TCF21过表达组、空载体组、未处理组。采用反转录PCR检测转染前后TCF21mRNA水平、Westernblot法检测TCF21蛋白水平；MTT法检测细胞的增殖能力，划痕实验检测转染后细胞的迁移能力；异硫氰酸荧光素标记的膜联素V／碘化丙啶(annexinV—FITC／PI)双标记结合流式细胞术检测细胞凋亡率，Westernblot法检测转染后KISSI、P53和基质金属蛋白酶9(MMP-9)的蛋白水平。结果TCF21基因转染SMMC-7721细胞后，细

3、胞内TCF21mRNA和蛋白水平均明显增加。与对照细胞相比，过表达TCF21的肝癌细胞增殖能力减弱、癌细胞迁移能力降低、细胞凋亡增加。Westernblot结果显示上调TCF21表达，可增加KISSI、P53水平并降低MMP-9的水平。结论过表达TCF21可抑制SMMC-7721肝癌细胞的增殖、迁移并促进其凋亡。[关键词]转录因子21(TCF21)；肝细胞肝癌；增殖；迁移；凋亡[中图分类号]R735．7，Q279，R392-33[文献标志码]AOver-expressionoftranscrip

4、tionfactor21inhibitstheproliferationandmigrationandpromotesapoptosisofSMMC-7721ceBsTANJingyu，ZHANGGuoqing，LIURui，ZHOUMi，LIZhongtang，WUZhongjunDepartmentofHepatobiliarySurgery，FirstAfiliatedHospital，ChongqingMedicalUniversity，Chongqing400016，ChinalAbs

5、tract]ObiectiveToexploretheimpactoftranscriptionfactor21(TCF21)geneonproliferation，migrationandapoptosisinSMMC-T／21hepatocelularcarcinomacellineandtherelatedmechanism．MethodsUsingLipofectamine~2000we，stablytransfectedpcDNA3．1(+)TCF21andpcDNA3．1(+)pla

6、smidsintoSMMC-7721cellsofTCF21over-expressiongroupandemptyvectorgroup，respectively．Theuntreatedcellsweresetasblankcontrolgroup．TheexpressionofTCF21mRNAandproteinwereinvestigatedbyreversetranscriptionPCRandWesternblotting．Celproliferationabilitywasdet

7、ectedbyM1_rassay．Woundhealingassaywasusedtoobservecellmigrationability．AnnexinV-FITC／Pldoublelabelingcombinedwithflowcytometrywasappliedtodeterminecellapoptosisrate．WesternblotingwasperformedtoexaminetheexpressionsofKISS1。P53andmatrixmetalloproteinas

8、e-9(MMP-9)．ResultsTCF21wasover-expressedinTCF'21-transfectedSMMC-7721cells．Comparedwiththecontrolgroups，theTCF2Iover-expressiongroupshowedrelativelyweakenedproliferationabilityandsignificantlyinhibitedmigrationabilityaswellastheincreasedapoptosis．Wes