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目的 :研究土贝母苷甲 (下称苷甲 )抗肿瘤作用的机制 ,探索它对细胞周期和细胞凋亡的影响。方法 :采用MTT法检测苷甲对细胞生长的抑制作用 ;采用流式细胞术 ,光学、荧光和电子显微镜 ,以及DNA琼脂糖凝胶电泳检查和分析苷甲对细胞周期和细胞凋亡的影响 ;采用蛋白免疫印迹法检测凋亡相关基因bcl 2和bax表达的变化。结果 :苷甲对HeLa细胞的生长有强抑制作用 ,其 2 4、4 8和 72h的IC50 值分别为 35 .7,2 3.6 ,17.4 μmol·L-1。在苷甲的作用下 ,细胞周期阻滞在G2 M期 ,细胞在形态学和生物化学上都出现了细胞凋亡的典型证据。蛋白免疫印迹结果揭示bcl 2下调 ,bax过表达。结论 :苷甲诱导的细胞周期阻滞和凋亡在苷甲的抗肿瘤效果中可能起着重要作用 ,而苷甲诱导的细胞凋亡与bcl 2的下调及bax的过表达密切相关。
Objective: To study the anti-tumor effect of timosaponin A and to explore its effect on cell cycle and apoptosis. Methods: MTT assay was used to detect the inhibitory effect of glycosides on cell growth. The effects of glycosides on cell cycle and apoptosis were examined by flow cytometry, optical, fluorescence and electron microscopy, and DNA agarose gel electrophoresis. Western blotting was used to detect the expression of apoptosis related genes bcl 2 and bax. RESULTS: Glycoside had a strong inhibitory effect on the growth of HeLa cells. IC50 values at 2, 4, 8 and 72 hours were 35.7, 23.6 and 17.4 μmol·L-1, respectively. Under the action of glycoside A, the cell cycle arrests at G2 M phase, and the cell morphologically and biochemically shows typical evidence of apoptosis. Western blot results revealed that bcl 2 was down-regulated and bax was overexpressed. Conclusion: Glycoside-induced cell cycle arrest and apoptosis may play an important role in the anti-tumor effect of glycoside A. Glycoside-induced apoptosis is closely related to the down-regulation of bcl-2 and bax overexpression.