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目的探讨Notch1基因沉默对人膀胱癌细胞增殖的影响。方法用pGenesil质粒构建靶向Notch1的siRNA真核表达载体psiRNA1,并将其转染到膀胱癌细胞系T24中,四甲基偶氮唑盐微量酶反应比色法(MTT)、流式细胞术(FCM)检测Notch1沉默后膀胱癌细胞生长变化、细胞周期和凋亡情况。逆转录聚合酶链反应(RT-PCR)和蛋白印记法(Western Blot)检测Notch1基因的mRNA和蛋白在转染前后表达变化。结果转染psiRNA1质粒后T24细胞的生长受到显著抑制,呈一定的时间依赖性(60 h内),凋亡率增加,G0/G1期细胞明显增多(未转染组、空载体组、阴性对照组),与对照组比较差异有统计学意义(P<0.05),且Notch1基因在mRNA和蛋白水平表达相对于对照组明显下调(P<0.05)。结论沉默Notch1基因能够抑制膀胱癌细胞的增殖,Notch1在膀胱癌中可能具有癌基因的作用。
Objective To investigate the effect of Notch1 gene silencing on human bladder cancer cell proliferation. Methods The eukaryotic expression vector psiRNA1 targeting Notch1 was constructed by pGenesil plasmid and transfected into bladder cancer cell line T24. Methyl thiazolyl tetrazolium assay (MTT), flow cytometry (FCM) to detect the growth of bladder cancer cells after Notch1 silencing, cell cycle and apoptosis. The expression of Notch1 mRNA and protein was detected by RT-PCR and Western Blot before and after transfection. Results After transfected with psiRNA1 plasmid, the growth of T24 cells was significantly inhibited (within 60 h), the apoptosis rate increased and the number of G0 / G1 phase cells significantly increased (untransfected group, empty vector group, negative control group (P <0.05). The expression of Notch1 mRNA and protein at mRNA and protein level was significantly lower than that of the control group (P <0.05). Conclusion Silencing Notch1 can inhibit the proliferation of bladder cancer cells. Notch1 may play an oncogene role in bladder cancer.