Apoptin基因诱导膀胱癌耐药细胞凋亡的研究

Acceleration of Apoptosis by Transfection of Apoptin Gene in Multi-drug Resistant Bladder Cancer Cells

目的观察Apoptin基因对膀胱癌耐药细胞株EJ/MDR细胞的杀伤效果,探讨其可能的机制。方法用脂质体将Apoptin基因导入EJ/MDR细胞,通过RT-PCR法检测Apoptin mRNA的表达,同时用SABC免疫组化法分析Apoptin和p53的表达。采用细胞计数法检测细胞生长抑制率,TUNEL法标记凋亡细胞,流式细胞仪检测细胞周期的变化。结果转入Apoptin基因后,EJ/MDR细胞的生长明显受抑制(P<0.05)。细胞周期分析可见凋亡峰,凋亡率35%。细胞中可见Apoptin阳性表达(P<0.05),p53表达无显著性差异(P>0.05)。凋亡细胞在荧光显微镜下形成凋亡小体。结论转入Apoptin基因可显著促进EJ/MDR细胞的凋亡,Apoptin诱导的凋亡不依赖功能性p53的生成。

Objective To observe the effects of Apoptin gene on killing MDR bladder cancer cells and illustrates its mechanisms. Methods Apoptin gene was transfected into MDR bladder cancer cells by liposome. The mRNA of Apoptin and bcl-2 were detected by RT-PCR. The protein of Apoptin and p53 were detected by SABC immunohistochemistry. The growth rate of human bladder cancer cells were studied by constructing the growth curve and TUNEL, cell apoptosis was observed by flow cytometry. Results The expression of Apoptin mRNA was positive in EJ/Apoptin cells （P〈0.05）. Apoptin protein expression of EJ/Apoptin cells was positive and p53 protein expression was not increased （P〉0.05）. The growth of MDR bladder cancer cells was significantly inhibited after bak gene was transfected （P〈0.05）. Apoptosis of cells increased significantly. The apoptosis rate was 35%. Conclusion Apoptin gene could inhibit the growth of MDR bladder cancer cells effectively. Inducing cell apoptosis by up-regulating the expression of p53 gene might not be one of its mechanisms.