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目的:研究重楼提取物单体重楼皂苷Ⅰ对膀胱癌细胞增殖和凋亡的影响。方法:体外培养膀胱尿路上皮细胞癌BIU-87细胞株,采用MTT法检测重楼皂苷Ⅰ对BIU-87细胞增殖的影响,计算半抑制率(IC50);采用流式细胞术检测重楼皂苷Ⅰ干预后BIU-87细胞凋亡的情况,透射电镜观察细胞超微结构的变化;采用实时定量PCR分析凋亡相关基因Bcl-2的变化,采用Western blot检测蛋白的表达变化。结果:重楼皂苷Ⅰ对BIU-87细胞的增殖有明显的抑制作用,24,48,72 h的IC50为9.71,5.91,4.68μmol·L-1。重楼皂苷Ⅰ对BIU-87细胞生长的抑制作用表现出时间和浓度依赖性。此外重楼皂苷Ⅰ干预以后,BIU-87细胞的凋亡率与对照组相比有显著性差异。实时定量PCR及Western blot的检测可见到凋亡相关基因Bcl-2的表达显著降低。结论:这些体外实验结果表明,重楼皂苷Ⅰ对膀胱癌有潜在的抗癌作用,下调Bcl-2基因的表达是其诱导膀胱癌细胞凋亡的作用机制之一。
OBJECTIVE: To study the effect of polysaccharide Saponin from Rhizoma Paridis extract on the proliferation and apoptosis of bladder cancer cells. Methods: The bladder transitional cell carcinoma BIU-87 cells were cultured in vitro. The proliferation of BIU-87 cells was detected by MTT assay and the half inhibitory rate (IC50) was calculated. The flow cytometry Ⅰ in vitro, the ultrastructural changes of BIU-87 cells were observed by transmission electron microscopy. The changes of apoptosis-related gene Bcl-2 were analyzed by real-time quantitative PCR. The protein expression was detected by Western blot. Results: Polygonum saponin Ⅰ significantly inhibited the proliferation of BIU-87 cells. The IC50 values at 24, 48 and 72 h were 9.71, 5.91 and 4.68 μmol·L -1, respectively. The inhibitory effect of polysaccharide Saponins Ⅰ on the growth of BIU-87 cells was time and concentration-dependent. In addition, after the intervention of polysaccharide saponins Ⅰ, the apoptosis rate of BIU-87 cells was significantly different from that of the control group. Real-time quantitative PCR and Western blot showed that the expression of apoptosis-related gene Bcl-2 was significantly decreased. CONCLUSION: These in vitro results indicate that Tail saponin Ⅰ has potential anticancer effect on bladder cancer, and down-regulation of Bcl-2 gene expression is one of the mechanisms of apoptosis induced by bladder cancer.