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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。
Objective: To study the effect and mechanism of octahedrus shikimic acid on apoptosis of human hepatocellular carcinoma cell line HepG-2. Methods: Human hepatocellular carcinoma HepG-2 cells in logarithmic growth phase were randomly divided into drug group and control group. HepG-2 cells were treated with different concentrations of 0.125, 0.5 and 1 g · L-1 octaketide for HepG-2 cells. MTT assay The effect of shikimic acid on the proliferation of HepG-2 cells was determined by flow cytometry. The effects of shikimic acid at the above concentrations on the cell cycle of HepG-2 cells were detected by immunocytochemistry. The apoptosis-related gene B-cell lymphoma leukemia 2 ( Bcl-2), Bcl-2 related X protein (Bax protein) expression changes. Results: The results of MTT assay showed that shikimic acid with various concentrations of 0.125, 0.5 and 1 g · L-1 significantly inhibited the growth of human hepatocellular carcinoma HepG-2 cells (P <0.05) in a time-and dose-dependent manner The results of flow cytometry showed that shikimic acid blocked the human hepatocellular carcinoma HepG-2 cells in G1 phase and prevented the cells from entering the S phase of DNA synthesis, thus inhibiting the proliferation of HepG-2 cells. Immunocytochemistry showed that the expression of Bcl-2 protein was significantly decreased, the expression of Bax was significantly increased, and the ratio of Bcl-2 / Bax was significantly decreased. Conclusions: Octagonal shikimic acid induced a large number of human hepatocellular carcinoma cell line HepG-2 to arrest in G1 phase in vitro, decreasing the number of cells entering S phase and inhibiting the cell proliferation. The effect of Octagonal shikimic acid may be induced by decreasing the ratio of Bcl-2 / Bax .