转染Livin基因对膀胱癌细胞凋亡的影响

目的 探讨凋亡抑制基因Livin对膀胱癌细胞凋亡的影响.方法 采用脂质体转染法将Livin基因转入膀胱癌T24细胞系,经G418筛选后获得稳定表达Livin的亚克隆细胞系T24/Livin+及仅转染载体的T24/pcDNA3.1(+).用丝裂霉素C(MMC)分别作用于转染前后的膀胱癌细胞系,应用四甲基偶氮唑盐(MTT)比色法检测转染前后细胞生长抑制率,应用流式细胞仪(FCM)及吖啶橙(AO)染色检测细胞凋亡.结果 成功建立了稳定表达Livin的亚克隆细胞系T24/Livin+;经MMC作用24 h后,T24/pcDNA3.1(+)细胞与T24的细胞凋亡率分别为(21.4±2.3)%和(19.6±2.3)%,而T24/Livin+的细胞凋亡率则为(8.7±1.5)%,差异有统计学意义(P＜0.01).结论 Livin基因提高了T24膀胱癌细胞的抗凋亡能力,特别是在膀胱癌化疗中显示出更强的抗凋亡能力.

Effects of gene Livin transfection affect on the apoptosis in bladder carcinoma cells

OBJECTIVE: To investigate the effect of gene Livin transfection on the apoptosis of human bladder transitional cell carcinoma (BTCC) cells. METHODS: Target fragment containing Livin full-length cDNA was obtained from the breast cancer cells of the line MCF7. Vector pcDNA3. 1 (+)-Livin was constructed and transfected into the bladder transitional cell carcinoma (BTCC) cells of the line T24 [T24/ Livin+ cells]. Another T24 cells were transfected with the eukaryotic expression vectors pcDNA3.1 (+) [T24/pcDNA3.1 (+) cells]. T24 cells without transfection were used as blank control group. These T24 cells underwent G418 selection so as to screen the T24/Livin+ cells stably expressing the target fragment. RT-PCR was used to detect the Livin expression level in the transfected cells. After the cells were treated with mitomycin C (MMC) for 24 h, MTT method was used to detect the inhibition rate. Acridine orange (AO) staining was used to observe the apoptotic cells. The apoptotic rate was observed by flow cytometry. RESULTS: RT-PCR results indicated that expression of Livin was positive in the T24/Livin+ cells, while were negative in the T24/pcDNA3.1 (+) and T24 cells. After being treated with MMC for 24 h, the apoptotic rate of the T24/Livin+ cell was (8.7 +/- 1.5)%, significantly lower than those of the T24/pcDNA3.1 (+) cells and T24 cells [(21.4 +/- 2.3)% and (19.6 +/- 2.3)% respectively, both P < 0.01]. CONCLUSION: The gene Livin increases the anti-apoptosis ability of tumor cells and the cell apoptosis induced by chemotherapy.