【摘要】 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。更多还原

【Abstract】 Objective To study the effect of miR-29 c on the apoptosis of prostate cancer cell line PC3. Methods The expression of miR-29 c,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-29 c overexpression adenovirus, the expression level of miR-29 c, 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 hoechst 33258 and FCM.Results The expression level of miR-29 c was lower and VEGF,VEGFR2 was higher in PC3 cell compared with that in RWPE-1( P<0. 001). The expression level of miR-29 c and BAX were higher and VEGF,p-VEGFR2 and Bcl-2 were lower in Ad-miR-29 c group compared with that in control group( P<0. 001).Higher apoptosis rate was detected in Ad-miR-29 c group compared with control group( P < 0. 001). Conclusions miR-29 c can promote apoptosis in prostate cancer cell PC3 by significantly inhibiting VEGF/VEGFR2 signaling.更多还原