TRPV2基因的RNAi对前列腺癌PC-3细胞增殖及侵袭能力的影响

Influence of TRPV2 silencing on prostate cancer cell PC-3 proliferation and migration

导出

摘要目的探讨瞬时感受器电位离子通道蛋白V亚族2(TRPV2)基因沉默对前列腺癌PC-3细胞增殖、细胞周期和侵袭能力的影响。方法构建针对TRPV2基因的小分子干扰RNA(siRNA)序列,在脂质体的介导下转染PC-3细胞;用RT-PCR测定TRPV2mRNA的表达;MTT法测定siRNA对PC-3细胞增殖的影响;流式细胞技术测定siRNA对PC-3细胞周期的影响;Tran-swell侵袭实验测定siRNA对PC-3细胞侵袭能力的影响。结果 TRPV2mRNA在激素非依赖PC-3细胞和DU145细胞中转录表达,而在激素依赖LNCaP细胞中未转录表达。靶向TRPV2的siRNA(siRNA-TRPV2)能有效地阻断PC-3细胞TRPV2基因在mRNA水平上的表达(P<0.01),且随着转染时间的延长而减少,转染72h后TRPV2mRNA的表达减少至对照组的15%。siRNA转染组穿过Matrigel膜的细胞数为(46.0±6.7),显著少于阴性对照组(92.0±9.4)和空白对照组(109.0±8.1)(P<0.01)。结论 TRPV2在激素非依赖前列腺癌细胞PC-3和DU145中转录表达。针对TRPV2的siRNA在体外能有效地抑制PC-3细胞株的TRPV2mRNA的转录,能显著抑制细胞的侵袭能力,但对细胞增殖和细胞周期没有显著影响。 Objective To explore the effects of TRPV2 gene silencing by small interfering RNA（siRNA）on the proliferatoin,cell cycle and migration of prostate cancer cell line PC-3.MethodssiRNA targeting TRPV2 gene was constructed and transfected into PC-3 cells mediated by liposome.Expression of TRPV2 mRNA was assayed by RT-PCR.The proliferation was detected by MTT method.The cell cycle was determined by FCM.The migration was detected by Transwell test.ResultsTRPV2 mRNA was expressed in androgen-independent PC-3 cells and DU-145,which was not expressed in androgen-dependent prostate cancer cell line LNCaP.Expression of TRPV2 mRNA in PC-3 cells was decreased in the extension of siRNA transfection.At 72h after transfection,the expression of TRPV2 mRNA was decreased by 85% compared with the blank and negative control group（P0.01）.The number of migration cell in siRNA transfection group was significantly less than that in blank group or negtive control group[（46.0±6.7）vs.（92.0±9.4）or（109.0±8.1）]（P0.01）.Conclusion TRPV2 is expressed in androgen-independent PC-3 cells and DU145 cells,which can be effectively inhibited by siRNA targeting TRPV2 gene.siRNA targeting TRPV2 gene has no effect on the cell proliferation and cell circle.