摘要
目的探讨蛇葡萄素在体外诱导人肝癌细胞株Bel-7402的凋亡作用及其可能机制。方法以不同浓度的蛇葡萄素作用于体外培养的人肝癌细胞株Bel-7402,应用MTT法检测培养24、48和72h的细胞生长情况;荧光染色观察细胞核的形态学改变;琼脂糖凝胶电泳分析细胞DNA变化;流式细胞仪检测细胞凋亡率;Western blot检测细胞caspase-3、Bcl-2和Bax的表达情况。结果蛇葡萄素对Bel-7402细胞的生长有明显的增殖抑制作用,呈浓度和时间依赖性。24、48、72h的IC50值分别是89.6±16.1、36.2±6.5和15.3±3.0mg·L-1。经荧光染色后可观察到典型的凋亡小体。细胞DNA电泳后呈现出凋亡细胞典型的DNAl adder。流式细胞术检测细胞凋亡率呈时间和浓度依赖性。caspase-3和Bcl-2蛋白表达量逐渐降低,Bax蛋白表达量逐渐增加,呈明显的浓度依赖性。结论蛇葡萄素能诱导体外培养的人肝癌Bel-7402细胞凋亡,其作用具有时间和浓度依赖性。下调Bcl-2、上调Bax表达,活化caspase-3是蛇葡萄素诱导Bel-7402细胞凋亡的可能机制之一。
Aim To investigate the effects of ampelopsin on induction of apoptosis in human hepatocellular carcinoma Bel-7402 cells.Methods Bel-7402 cells were treated with ampelopsin with different concentrations for 24,48 and 72 h.The cell proliferation was detected by MTT assay.The morphological change of cells was observed through microscope observation by fluorescence staining.DNA fragmentation was visualized by agarose gel electrophoresis.The apoptosis rate was analyzed by flow cytometry.The expressions of caspase-3,Bcl-2 and Bax protein were detected by Western blot.Results Ampelopsin inhibited the proliferation of Bel-7402 cell line in a dose-and time-dependent manner.The IC50 values were 89.6±16.1,36.2±6.5 and 15.3±3.0 mg·L-1 at 24,48 and 72 h,respectively.The fluorescence microscope showed clearly cell apoptosis with apoptotic body.Agarose gel electrophoresis result showed that Bel-7402 treated with ampelopsin produced a DNA ladder band.The sub-G1 peak was detected and resulted in dose-and time-dependent increasing of the population of sub-G1 DNA content by flow cytometry.The expression of Bcl-2 protein was down-regulated,while the expression of Bax protein was up-regulated.The pro-caspase-3 protein was down-expressed and activated.Conclusions Ampelopsin could inhibit Bel-7402 proliferation through inducing cell apoptosis.The mechanism might be that ampelopsin could directly or indirectly enhance the level of anti-apoptosis protein Bcl-2 and decrease the level of apoptosis protein Bax.The pathway of pro-caspase-3 activated was initiated and effector caspase-3 was sequentially activated.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2009年第11期1502-1506,共5页
Chinese Pharmacological Bulletin
基金
广东省科技计划资助项目(No2003C104033)
