Objective To study the effect of miR-29c on the apoptosis of prostate cancer cell line PC 3.Methods The expression of miR-29c, VEGF and VEGFR2 in RWPE-1 and PC3 were detected to verify the different between normal and cancer cell .The location of p-VEGFR2 was measured by immunofluorescence (IF).After PC3 cell were transfected by miR-29c overexpression adenovirus , the expression level of miR-29c, VEGF, t-VEGFR2, p-VEGFR2, BAX and Bcl-2 were measured by RT-PCR and Western blot , the cell apoptosis rate was detected by the kits of ho-echst 33258 and FCM.Results The expression level of miR-29c was lower and VEGF, VEGFR2 was higher in PC3 cell compared with that in RWPE-1(P<0.001).The expression level of miR-29c and BAX were higher and VEGF , p-VEGFR2 and Bcl-2 were lower in Ad-miR-29c group compared with that in control group (P<0.001).Higher apop-tosis rate was detected in Ad-miR-29c group compared with control group (P<0.001).Conclusions miR-29c can promote apoptosis in prostate cancer cell PC 3 by significantly inhibiting VEGF/VEGFR2 signaling.%目的 研究miR-29c对前列腺癌PC3细胞凋亡的影响及其机制.方法 以人正常前列腺上皮细胞系(RWPE-1)为对照,检测前列腺癌PC3细胞系中miR-29c、血管内皮生长因子(VEGF)及血管内皮生长因子受体2(VEGFR2)的表达;免疫荧光(IF)检测p-VEGFR2在细胞中的定位;miR-29c过表达腺病毒感染PC3,real-time PCR检测miR-29c及VEGF的表达;Western blot检测VEGF、t-VEGFR2、p-VEGFR2、BAX和Bcl-2的表达;hoechst 33258染色法检测PC3细胞凋亡;流式细胞计量术(FCM)检测PC3细胞早期凋亡率.结果 与RWPE-1相比,PC3细胞miR-29c表达降低,VEGF及VEGFR2表达升高(P<0.001);与对照组相比,miR-29c过表达组VEGF、p-VEGFR2及Bcl-2的表达显著降低(P<0.001),BAX的表达显著升高(P<0.001),细胞凋亡与早期凋亡率显著增加(P<0.001).结论 重新表达miR-29c可以显著抑制VEGF/VEGFR2信号通路并促进PC3细胞凋亡.
展开▼
