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目的:观察凋亡素对表达Survivin膀胱癌细胞株的作用效果,并结合相应机制探讨其临床意义。方法:利用免疫组织化学法检测膀胱癌细胞株BIU-87中Survivin的表达水平,并计算平均每高倍镜视野中阳性细胞比率。将肿瘤细胞分为单纯细胞组、转染空质粒pCDNA3组及转染pCDNA3/Apoptin组,利用脂质体转染法将真核表达载体pCDNA3/Apoptin及pCDNA3空质粒转染入膀胱癌细胞中,并于转染后24h通过RT-PCR法检测细胞中Apoptin的表达情况。分别于转染后24、48、72h利用MTT法检测各组细胞存活率,并利用流式细胞仪TUNEL法检测各组细胞的凋亡率。结果:利用免疫组织化学法检测发现膀胱癌细胞株BIU-87中Survivin呈高表达状态,平均每个高倍镜视野中阳性细胞比例约70%,并有较多细胞呈现高表达状态。转染后24h通过RT-PCR法可见Apoptin在膀胱癌细胞中表达。分别于转染后24、48、72h利用MTT法检测各组细胞存活率,发现转染pCDNA3/Apoptin组肿瘤细胞存活率均明显低于对照组,差别有统计学意义(P<0.05);利用流式细胞仪TUNEL法检测各组细胞的凋亡率,发现转染pCDNA3/Apoptin组肿瘤细胞凋亡率均明显高于对照组,差别有统计学意义(P<0.05)。结论:凋亡素可在体外诱导高表达Survivin的膀胱癌细胞株BIU-87高效凋亡,结合相关机制表明凋亡素可在一定程度上克服Survivin的抗凋亡作用,因此有可能成为Survivin拮抗的抗肿瘤药物的协同用药选择。
Objective: To observe the effect of apoptin on the expression of Survivin in bladder cancer cell lines, and to explore its clinical significance with the corresponding mechanism. Methods: The expression of Survivin in bladder cancer cell line BIU-87 was detected by immunohistochemistry and the ratio of positive cells in each high magnification field was calculated. The tumor cells were divided into simple cell group, transfected with empty plasmid pCDNA3 group and transfected pCDNA3 / Apoptin group, transfected into bladder cancer cells by liposome transfection method, the eukaryotic expression vector pCDNA3 / Apoptin and pCDNA3 empty plasmid, 24h after transfection, the expression of Apoptin in cells was detected by RT-PCR. The cell viability was measured by MTT assay at 24, 48, 72 h after transfection, and the apoptosis rate of each group was detected by flow cytometry TUNEL method. Results: Survivin was highly expressed in bladder cancer cell line BIU-87 detected by immunohistochemical staining. The average percentage of positive cells in each high magnification field was about 70%, and more cells were highly expressed. Apoptin was expressed in bladder cancer cells by RT-PCR 24h after transfection. The survival rates of all the groups were detected by MTT assay at 24, 48 and 72h after transfection. The survival rate of tumor cells transfected with pCDNA3 / Apoptin was significantly lower than that of the control group (P <0.05) Flow cytometry TUNEL assay of apoptosis rate of each group of cells found that the transfected pCDNA3 / Apoptin tumor cell apoptosis rate were significantly higher than the control group, the difference was statistically significant (P <0.05). Conclusion: Apoptosis can induce apoptosis of bladder cancer cell line BIU-87 with high expression of Survivin in vitro. The combination of these mechanisms suggests that apoptin can overcome the anti-apoptotic effect of Survivin to a certain extent and thus may be a Survivin antagonist Antineoplastic drugs synergistic choice.