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目的:研究红芪多糖对人肝癌HEP-G2细胞的增殖抑制与诱导凋亡作用及其可能机制。方法:应用MTT法、显微荧光术和流式细胞术测定红芪多糖对HEP-G2细胞的增殖、细胞周期及凋亡的影响,并应用流式细胞术测定其对Bc1-2和Bax蛋白表达的影响。结果:红芪多糖对HEP-G2细胞的增殖抑制作用具有时间及剂量依赖性,400μg/mL红芪多糖作用72 h后对HEP-G2细胞抑制率达到33.8%;显微荧光术发现作用72 h后,HEP-G2细胞核明显固缩、凝聚,出现浓染致密的颗粒块状荧光;流式细胞术检测显示200、400μg/mL红芪多糖作用72 h,可以观察到G0前期的亚二倍体DNA峰,但对G0/G1期细胞比例和S期细胞比例无明显影响,而G2/M期细胞比例显著升高(P<0.05);Bc1-2蛋白表达水平降低,Bax蛋白表达水平提高。结论:红芪多糖可通过抑制人肝癌HEP-G2细胞的生长增殖、G2/M期阻滞、诱导细胞凋亡、降低bc l-2蛋白、提高Bax蛋白表达等作用机制发挥抗肿瘤作用。
OBJECTIVE: To study the effect of Radix Hedysari Polysaccarides on proliferation inhibition and apoptosis of human hepatocellular carcinoma HEP-G2 cells and its possible mechanism. METHODS: MTT assay, micro-fluorescence, and flow cytometry were used to determine the effects of HRP on proliferation, cell cycle, and apoptosis of HEP-G2 cells. Flow cytometry was used to determine Bcl-2 and Bax proteins. The effect of expression. RESULTS: The proliferation inhibition of HEP-G2 cells was inhibited in a time and dose-dependent manner. The inhibition rate of HEP-G2 cells reached to 33.8% after 400 μg/mL HA treatment for 72 h; the effect was found after 72 h of microfluorescence. Afterwards, the HEP-G2 cell nuclei were significantly pyknosis and agglutination, and dense and dense granular fluorescence appeared. Flow cytometry showed that the effect of 200, 400 μg/mL HRP could be observed for 72 h, and G0 prophase hypodiploidy could be observed. The peak of DNA had no significant effect on the proportion of G0/G1 phase cells and the proportion of S phase cells, but the proportion of cells in G2/M phase increased significantly (P<0.05); the expression of Bc1-2 protein decreased and the expression of Bax protein increased. Conclusion: HRP could exert its anti-tumor effect by inhibiting the growth and proliferation of human liver cancer HEP-G2 cells, blocking G2/M phase, inducing cell apoptosis, decreasing bcl-2 protein and increasing Bax protein expression.