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目的研究人参皂甙Rh2(GSRh2)对肝癌Bel7404细胞增殖和凋亡的影响及其可能的机制。方法采用MTT比色法观察GSRh2对体外培养的Bel7404细胞生长的抑制作用;应用流式细胞仪检测凋亡细胞和细胞周期变化;采用免疫组化SP法检测药物处理前后凋亡相关基因产物p53、Bax和Bcl2蛋白表达。结果①MTT比色法测定显示,Bel7404细胞经GSRh2作用后生长受抑制,呈剂量依赖性和时间依赖性,半数抑制浓度(IC50)为27.6μgml。②流式细胞仪检测证实,GSRh2能诱导Bel7404细胞凋亡。③当GSRh2浓度由12.5μgml增加至50.0μgml时,细胞凋亡率(AR)依次升高;但GSRh2浓度进一步增加时,AR未见相应升高。④GSRh2可将细胞周期阻滞于G1期。⑤GSRh2处理细胞后,突变型p53表达下调而Bax表达上调,Bcl2在GSRh2处理前后无明显变化。结论①GSRh2能抑制体外培养的Bel7404细胞增殖并诱导其凋亡;②GSRh2诱导Bel7404细胞凋亡可能是通过上调Bax表达、下调突变型p53表达以及阻滞细胞周期而实现。
Objective To investigate the effect of ginsenoside Rh2 (GSRh2) on proliferation and apoptosis of human hepatoma Bel7404 cells and its possible mechanism. Methods MTT colorimetric assay was used to observe the inhibitory effect of GSRh2 on the growth of Bel7404 cells in vitro. Flow cytometry was used to detect apoptotic cells and cell cycle changes. Immunohistochemical SP method was used to detect the expression of p53, Bax and Bcl2 protein expression. Results ① MTT colorimetric assay showed that the growth of Bel7404 cells was inhibited by GSRh2 in a dose-dependent and time-dependent manner. The IC50 was 27.6μgml. ② Flow cytometry confirmed that GSRh2 can induce Bel7404 cell apoptosis. ③ When the concentration of GSRh2 was increased from 12.5μgml to 50.0μgml, the apoptotic rate (AR) increased in turn; however, when GSRh2 concentration increased further, AR did not increase correspondingly. ④ GSRh2 can block the cell cycle in G1 phase. ⑤ GSRh2 treated cells, the mutant p53 expression was down-regulated Bax expression, Bcl2 GSRh2 treatment before and after no significant change. Conclusion ① GSRh2 can inhibit the proliferation and induce the apoptosis of Bel7404 cells cultured in vitro. ② The apoptosis induced by GSRh2 may be related to up-regulating the expression of Bax, down-regulating the expression of mutant p53 and arresting the cell cycle.