tcf4对人皮肤鳞状细胞癌a431细胞株生物学行为的影响

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1、第36卷第4期第三军医大学学报V0l_36．No．43602014年2月28日JThirdMilMedUnivJan．282014文章编号：1000-5404(2014)04-0360—04TCF4对人皮肤鳞状细胞癌A431细胞株生物学行为的影响刘燕，熊亚，宋志强，沈柱，郝飞，杨希川(400038重庆，第三军医大学西南医院皮肤科，重庆市皮肤性病研究所)[摘要]目的探讨T细胞因子4(Tcellfactor4，TCF4)小干扰RNA(siRNA)对人皮肤鳞状细胞癌A431细胞株TCF4基因表达的抑制作用及对细胞增殖、迁移、侵袭及周期的影响。方法转染TCF4siRNA至A4

2、31细胞株。应用RT．PCR检测A431细胞TCF4mRNA的表达，Westernblot检测TCF4蛋白的表达，CCK一8方法检测上述各组细胞增殖率(1、2、3、4d及5d)，小室迁移实验检测细胞迁移情况，Transwell实验检测侵袭情况，流式细胞仪检测细胞周期(24h)。结果与未转染A431细胞株和转染空载体的A43l细胞株比较，转染TCF4siRNA表达载体1、2、3、4d及5d的细胞增殖率明显降低(P<0．05)；TCF4siRNA表达载体组细胞迁移和侵袭能力明显下降(P<0．05)；细胞周期分析发现转染TCF4siRNA使得A431细胞株出现明显的G：／M

3、期阻滞现象。结论采用TCF4特异性siRNA能够抑制皮肤鳞状细胞癌TCF4的表达，从而抑制皮肤鳞状细胞癌的增殖。[关键词]TCF4；鳞状细胞癌；增殖；凋亡；Wnt／[3一catenin信号通路[中图法分类号]R394．3；R730．23；R739．5[文献标志码]AEffectofTCF4onbiologicalbehaviorofhumansquamouscarcinomaA431cellsLiuYan，XiongYa，SongZhiqiang，ShenZhu，HaoFei，YangXichuan(DepartmentofDermatology，ChongqingI

4、nstituteofDermatovenereology，SouthwestHospital，ThirdMilitaryMedicalUniversity，Chongqing，400038，China)[Abstract]ObjectiveToinvestigatetheeffectsofTcellfactor4(TCF4)siRNAonTcF4geneexpression，andthemigration，invasionandcellcycleofhumansquamouscarcinomaA431cells．MethodsA43lcellsweretransf~c

5、tedwithTCF4siRNAliposome．ThemRNAexpressionofTCF4inA431cellswasde—tectedbyRT．PCR．TheexpressionofTCF4proteinwasdetectedbyWesternblotting．Theproliferation．migra-tion．andinvasionactivitiesandcellcycleofA431cellsweremeasuredbyCCK8assay，chambercellmigra—tionassay．transwellchambertestandflowcy

6、tometry，respectively．ResultsTCF4siRNAtransfectioncouldinhibitsignificantlytheexpressionofTCF4mRNAandproteininA431cells．TheratesofcellproliferationandabilitiesofmigrationandinvasionwerealsosignificantlydecreasedafterTCF4siRNAtransfection．Inaddition．analysisofcellcycledemonstratedamarkedG

7、／MarrestoftheA43lcellstreatedwithTCF4siRNA．ConclusionItiSconcludedthatTCF4siRNAcaninhibitproliferationofhumansquamouscarcinomacells．whichmayprovidearationalapproachinclinicaltherapyforhumansquamouscarcinoma．[Keywords]TcF4；humansquamouscarcinoma；proliferation；apoptosis；Wnt／~-eat