摘要
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 examine the effect of ginsenoside Rh 2(GS Rh 2) on proliferation and apoptosis of Bel 7404 human hepatoma cell line,and to explore its potential action mechanism.Methods Cell growth inhibition was evaluated by MTT colormetric assay.Apoptotic cells and the change of cell cycle were determined by flow cytometry (FCM).Furthermore,the proteins produced by apoptosis related gene such as p53,Bax and Bcl 2 were detected with immunohistochemistry method before and after GS Rh 2 treatment.Results ①MTT colormetric assay revealed that the inhibition of GS Rh 2 to the growth of Bel 7404 cells was in a time and dose dependent manner,the 50% inhibition concentration (IC 50 ) value was 27.6 μg/ml.②GS Rh 2 could induce apoptosis on Bel 7404 cells as indicated by FCM.③When the concentration of GS Rh 2 was increased from 12.5 μg/ml to 50.0 μg/ml,the apoptosis rate (AR) of Bel 7404 cell line elevated from 16.2% to 27.5% accordingly;but when the concentration of GS Rh 2 was increased in further,the AR no longer rose.④GS Rh 2 was capable of arresting the cell cycle at G 1 phase.⑤The expression of mutated p53 protein was down regulated and that of Bax was up regulated after exposure to GS Rh 2,but the expression of Bcl 2 protein did not change significantly.Conclusions ①GS Rh 2 exhibited growth inhibition effect and induced apoptosis on Bel 7404 cell line in vitro .②Apoptosis of Bel 7404 cells induced by GS Rh 2 may be related with arresting cell cycle,up regulating Bax protein expression and down regulating mutated p53 protein expression.
出处
《中国肿瘤临床与康复》
2004年第4期289-292,共4页
Chinese Journal of Clinical Oncology and Rehabilitation
关键词
人参皂甙-Rh2
肝肿瘤
增殖
凋亡
GS-Rh_2(ginsenoside-Rh_2)
Liver neoplasm
Proliferation
Apoptosis
