美味猕猴桃根乙酸乙酯提取物部分对肝癌HepG2细胞增殖影响及机制研究

Effect of Actinidia Deliciosa extracted by ethylacetate proliferation of HepG2 cells and its mechanism

目的探讨美味猕猴桃根乙酸乙酯提取物部分(EE-ADP)对肝癌HepG2细胞增殖的影响及其机制。方法四甲基偶氮唑蓝(MTT)检测EE-ADP对肝癌HepG2细胞增殖的作用;Hochest33258染色及流式细胞技术检测其凋亡作用;蛋白免疫印迹法检测EE-ADP对抗原呈递细胞(APC)蛋白及β连环蛋白(β-catenin)的影响及可能影响细胞增殖的机制。结果 MTT染色法提示,不同浓度的EE-ADP干预肝癌HepG2细胞能一定程度上抑制肝癌细胞的增殖,且存在时间-浓度依赖性。40μg/ml组的凋亡率为17. 70%,与完全培养基组的4. 80%比较,差异无统计学意义(P> 0. 05)。60μg/ml组及80μg/ml组凋亡率分别为50. 75%、64. 30%,与完全培养基组比较,差异均有统计学意义(P <0. 05)。蛋白免疫印迹法结果显示,随着加药浓度的增加,APC基因蛋白表达量增加,β-catenin基因表达量减少。结论 EE-ADP可以抑制肝癌HepG2细胞的增值,其机制可能为降低β-catenin而上调APC的表达,阻断Wnt/β-catenin信号通路。

Objective To investigate the effect of actinidia deliciosa planch extracted by ethyl acetate( EE-ADP) on HepG2 cell proliferation and its possible mechanism. Methods The effect of EE-ADP on HepG2 cell proliferation was detected by MTT assay. Hochest33258 staining and flow cytometry were used to detect the apoptotic proteins. Western blot was used to detect the effect of EEADP on antigen presenting cell( APC) protein and β-catenin and the possible mechanism of affecting cell proliferation. Results MTT staining suggested that EE-ADP intervention in HCC HepG2 cells at different concentrations could inhibit the proliferation of HCC cells to a certain extent,and there was a time-concentration dependence. The apoptosis rate of 40 μg/ml group was 17. 70%,compared with4. 80% of the complete medium group,showing no significant difference( P > 0. 05). The apoptosis rates of 60 μg/ml and 80 μg/ml groups were 50. 75% and 64. 30%,respectively,with statistically significant differences compared with the complete medium group( P < 0. 05). The results of Western blot showed that with the increase of concentration,the protein expression of APC increased and the expression of β-catenin reduced. Conclusion EE-ADP can inhibit the proliferation of hepatocellular carcinoma HepG2 cells,and the mechanism may be that the up-regulated APC expression blocks the Wnt/β-catenin signaling pathway to reduce β-catenin.