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目的:以人肝癌HepG2细胞为研究对象,探讨phaseoloideside E(PE)的体外抗肿瘤活性。方法:采用MTT法,检测不同浓度的PE处理48 h后对HepG2细胞的细胞毒性效应;应用光学显微镜、荧光显微镜观察PE处理下的细胞形态学变化;用DNA琼脂糖凝胶电泳技术检测凋亡,并用免疫印迹法检测PE对凋亡相关蛋白Bax,Bcl-2表达水平的影响。结果:PE对HepG2细胞的生长有明显的细胞毒性效应;直接形态显微观察及Hoechst 33258荧光染色皆可见典型的凋亡细胞形态学特征;琼脂糖凝胶电泳结果显示给药组出现明显的DNA梯状凋亡带,而对照组没有出现梯状条带;免疫印迹结果显示PE可以增加Bax表达并减少Bcl-2表达。结论:PE能够诱导人肝癌HepG2细胞凋亡,并提示PE诱导的Bax,Bcl-2蛋白的表达水平变化可能与它的凋亡诱导效应紧密相关。
Objective: To investigate the antitumor activity of phaseoloideside E (PE) in vitro in human hepatocellular carcinoma HepG2 cells. Methods: MTT assay was used to examine the cytotoxicity of HepG2 cells treated with different concentrations of PE for 48 h. The morphological changes of cells treated with PE were observed with light microscope and fluorescence microscope. Apoptosis was detected by DNA agarose gel electrophoresis , And Western blotting was used to detect the effect of PE on the expression of Bax and Bcl-2. Results: PE had obvious cytotoxity effect on HepG2 cells. Morphological characteristics of apoptotic cells were observed by direct morphology microscopy and Hoechst 33258 fluorescence staining. The results of agarose gel electrophoresis showed that there was obvious DNA Ladder-like apoptosis zone, while the control group did not appear ladder-like bands; Western blot results showed that PE can increase Bax expression and reduce Bcl-2 expression. CONCLUSION: PE can induce apoptosis of HepG2 cells and suggest that the changes of Bax and Bcl-2 protein expression induced by PE might be closely related to its apoptosis-inducing effect.