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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 investigate the effect of transient receptor potential ion channel protein subunit 2 (TRPV2) gene silencing on the proliferation, cell cycle and invasion of prostate cancer PC-3 cells. Methods The small interfering RNA (siRNA) sequence targeting TRPV2 gene was constructed and transfected into PC-3 cells by lipofectamine. The expression of TRPV2 mRNA was detected by RT-PCR. The proliferation of PC-3 cells was detected by MTT assay The influence of siRNA on the cell cycle of PC-3 cells was determined by flow cytometry. The effect of siRNA on the invasiveness of PC-3 cells was evaluated by Tran-swell invasion assay. Results TRPV2 mRNA was transcriptally expressed in hormone-independent PC-3 cells and DU145 cells, but not in hormone-dependent LNCaP cells. The siRNA targeting TRPV2 (siRNA-TRPV2) effectively blocked the expression of TRPV2 mRNA in PC-3 cells (P <0.01), and decreased with the prolongation of transfection time. After 72 h of transfection, TRPV2 mRNA The expression was reduced to 15% of the control group. The number of cells passing through the Matrigel membrane in the siRNA transfection group was (46.0 ± 6.7), significantly lower than that in the negative control group (92.0 ± 9.4) and the blank control group (109.0 ± 8.1) (P <0.01). Conclusion TRPV2 is transcribed in hormone-independent prostate cancer cell lines PC-3 and DU145. SiRNA targeting TRPV2 effectively inhibited TRPV2 mRNA transcription in PC-3 cells in vitro and significantly inhibited cell invasion but had no significant effect on cell proliferation and cell cycle.