靶向沉默Notch1基因对人膀胱癌细胞生物学行为的影响

目的 探讨沉默Notch1基因对人膀胱癌细胞生物学行为的影响. 方法 将靶向Notch1的siRNA真核表达载体psiRNA Notch1-1转染膀胱癌细胞株T24和BIU-87,噻唑盐法、流式细胞术检测沉默Notch1后膀胱癌细胞生长、细胞周期和凋亡情况.RT-PCR和蛋白质印迹法检测转染前后Notch1基因mRNA和蛋白表达的变化. 结果 转染后72 h,T24和BIU-87细胞G0/G1期细胞比例明显增加,分别为(80.13±2.69)%和(69.44±2.41)%,与T24对照组(23.89±1.32)%和BIU-87对照组(24.63±1.68)%比较差异有统计学意义(P值均<0.05).转染后72 h,T24和BIU-87细胞凋亡率分别为(13.75±1.23)%和(8.72±1.01)%,与T24对照组(1.28±0.14)%和BIU-87对照组(1.01±0.27)%比较差异均有统计学意义(P值均<0.05).转染后24 h细胞生长明显受到抑制,该抑制作用持续到转染后96 h.转染后72 h,T24和BIU-87细胞中Notch1基因mRNA和蛋白表达明显下调(P<0.05). 结论 Notch1在膀胱癌细胞株中可能起致癌作用.沉默Notch1基因能够抑制膀胱癌细胞的增殖.

Effect of Notch1 on biological behavior of bladder cancer cells

Objective To investigate the in vitro effects of bladder cancer cell proliferation after silencing Notch1 gene. Methods The siRNA eukaryotic expression vector of Notch1 (psiRNA1)was constructed and transfected into bladder cancer cell lines T24 and BIU-87. Methabensthiazuron (MTT) and flow cytometry (FCM) assays were used to detect bladder cancer cells line growth, cell cycle and apoptosis after the transfection. RT-PCR and Western blotting were used to determine the expression changes of Notch1 in these cell lines. Results After transfection for 72 h, the rate of G0/G1 phase cells inceased from (23.89±1.32) % to (80.13±2.69)% in T24 cell line, and increased from (24.63±1.68)% to (69.44±2.41)% in BIU-87 cell line (both P<0.05). In addition, apop-totic cell index in T24 and BIU-87 cell lines increased from (1.28±0.14)% to (13.75±1.23)%, from (1.01±0.27)% to (8.72±1.01)%, respectively(both P<0.05). The growth of T24 and BIU-87 cell lines was obviously inhibited 24 h after the transfection, and the inhibitory effects lasted until 96 h after the transfection. Notch1 mRNA and protein significantly downregulated after transfection compared to the control(P<0.05). Conclusions Silencing Notch1 expression can inhibit the prolif-eration of bladder cancer cell lines. Notch1 gene might act as a tumor gene in bladder cancer.