摘要
目的探讨沉默Notchl基因对人膀胱癌细胞生物学行为的影响。方法将靶向Notchl的siRNA真核表达载体psiRNANotchl-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Notchl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Notchl基因mRNA和蛋白表达的变化。结果转染后72h．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72h，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BIU87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24h细胞生长明显受到抑制，该抑制作用持续到转染后96h。转染后72h，T24和BIU87细胞中Notchl基因mRNA和蛋白表达明显下调（P〈0．05）。结论Notchl在膀胱癌细胞株中可能起致癌作用，沉默Notchl基因能够抑制膀胱癌细胞的增殖。
Objective To investigate the in vitro effects of bladder cancer cell proliferation after silencing Notchl gene. Methods The siRNA eukaryotic expression vector of Notchl（psiRNAl）was constructed and transfeeted into bladder cancer cell lines T24 and BIU-87. Methabenzthiazuron （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 Notchl 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 BIU87 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. Notchl mRNA and protein significantly downregulated after transfection compared to the control（P〈0.05）. Conclusions Silencing Notchl expression can inhibit the proliferation of bladder cancer cell lines. Notchl gene might act as a tumor gene in bladder cancer.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
2009年第5期328-331,共4页
Chinese Journal of Urology
基金
基金项目：国家自然科学基金资助项目（30571858）
