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目的:探讨甘露聚糖肽联合5-氟脲嘧啶(5-FU)对人肝癌HepG2细胞增殖及凋亡的影响及其作用机制。方法:将HepG2细胞分为对照组、5-FU组、甘露聚糖肽组及联合用药组。利用MTT法观察药物对各组细胞增殖的影响。流式细胞术检测各组细胞的周期时相变化、增殖指数(PI)及凋亡率。免疫细胞化学法分析各组细胞Bcl-2蛋白的表达情况。结果:甘露聚糖肽单独应用后对HepG2细胞的增殖抑制作用较弱,与5-FU联合使用后能明显增强对细胞生长的抑制作用,并呈时间和剂量依赖性。药物作用细胞48h后,与对照组及5-FU组相比,联合用药组G_0/G_1期细胞比率增加,细胞凋亡率明显增高(P<0.01)。免疫细胞化学法分析显示,两药联合使用可使细胞Bcl-2蛋白的表达明显下降(P<0.01)。结论:甘露聚糖肽与5-FU联合应用具有抑制HepG2细胞的增殖及诱导其凋亡的作用,其机制之一可能与下调Bcl-2蛋白的表达相关。
Objective: To investigate the effect and mechanism of mannan oligopeptide combined with 5-fluorouracil (5-FU) on proliferation and apoptosis of human hepatocellular carcinoma HepG2 cells. Methods: HepG2 cells were divided into control group, 5-FU group, mannan group and combination group. MTT assay was used to observe the effects of drugs on cell proliferation in each group. Flow cytometry was used to detect the cell cycle changes, proliferation index (PI) and apoptosis rate. The expression of Bcl-2 protein in each group was analyzed by immunocytochemistry. Results: Mannatide alone had a weak inhibitory effect on the proliferation of HepG2 cells. Combined with 5-FU, the inhibitory effect of Mannatide on HepG2 cells could be obviously enhanced in a time-and dose-dependent manner. Compared with the control group and the 5-FU group, the percentage of G 0 / G 1 phase cells in the combination group increased and the apoptosis rate increased significantly (P <0.01) 48 h after the drug-treated cells were treated. Immunocytochemistry analysis showed that the combination of the two drugs can significantly reduce the expression of Bcl-2 protein (P <0.01). Conclusion: The combination of mannan peptide and 5-FU can inhibit the proliferation and induce the apoptosis of HepG2 cells. One of the mechanisms may be related to the down-regulation of Bcl-2 protein expression.