八角莽草酸诱导人肝癌细胞HepG-2凋亡及其机制研究

Shikimic Acid Inducing Apoptosis on Hepatocellular Carcinoma HepG-2 Cells Through Decreasing the Ratio of Bcl-2/Bax

摘要:

目的：研究八角莽草酸诱导人肝癌细胞HepG-2凋亡的作用和机制。方法：取对数生长期人肝癌HepG-2细胞，随机分为药物组和对照组，用0.125，0.5，1 g·L^-1不同浓度八角莽草酸作用人肝癌HepG-2细胞，采用MTT法测定莽草酸对HepG-2细胞增殖的影响；通过流式细胞仪检测在以上浓度的莽草酸对肝癌HepG-2细胞周期的影响；通过免疫细胞化学检测凋亡相关基因B细胞淋巴瘤白血病2（Bcl-2）、Bcl-2相关X蛋白（Bax蛋白）表达的改变。结果：MTT实验结果表明，各质量浓度0.125，0.5，1 g·L^-1的莽草酸对人肝癌HepG-2的生长具有显著的抑制作用（P<0.05），呈现时间依赖性和剂量依赖性；流式结果表明莽草酸将人肝癌HepG-2细胞阻滞在G₁期，使细胞不能顺利地进入细胞DNA合成期S期，从而抑制了该细胞的增殖。免疫细胞化学检测凋亡相关基因Bcl-2蛋白表达明显减少、Bax表达明显增加；Bcl-2/Bax比例明显降低。结论：八角莽草酸在体外诱导人肝癌HepG-2细胞大量阻滞在G₁期，进入S期细胞减少，抑制细胞增殖，其作用可能通过降低Bcl-2/Bax比例来诱导细胞凋亡而实现的。

Abstract:

Objective: To investigate the effects of shikimic acid on inducing hepatocellular carcinoma HepG-2 cells apoptosis in vitro. Method: The MTT assay was used, and the hepatocellular carcinoma HepG-2 cells in logarithmic phase of growth were randomly divided into control group and treatment group treated with the concentration of 0.125, 0.5, 1 g·L^-1 shikimic acid in hepatocellular carcinoma HepG-2 cells. The cell cycle distribution was analyzed by flow cytometry. The protein expression of B cell lymphoma/leukemia-2(Bcl-2) and Bcl-2 associated x protein(Bax) was analyzed by immunocytochemical method. Result: Shikimic acid inhibited the growth of cells significantly(P<0.05). The suppression was both in a time-and dose-dependent manner. The flow cytometry showed that shikimic acid can inhibit the proliferation by preventing cells turning into S phase from G₁ phase in cell cycle, at the same time, shikimic acid down-regulated the Bcl-2 expression and up-regulated the Bax expression, the ratio of Bcl-2/Bax was evident decrease. Conclusion: Shikimic acid can inhibit proliferation hepatocellular carcinoma HepG-2 cells and induce a G₁ cell cycle arrest and reduce the cell population in S phase, it may induce apoptosis through decreasing the ratio of Bcl-2/Bax.