镰形棘豆诱导人肝癌细胞SMMC-7721凋亡及初步的机制研究

Study on apoptosis of SMMC-7721 hepatocarcinoma cells induced by Oxytropis falcata and its preliminary mechanism

目的:观察镰形棘豆总黄酮对人肝癌细胞SMMC-7721增殖的影响及初步的机制探讨。方法:不同剂量的镰形棘豆总黄酮处理SMMC-7721细胞24 h后,MTT法观察总黄酮对细胞增殖的抑制作用;应用光学和荧光显微镜观察总黄酮对SMMC-7721细胞形态的影响;应用流式细胞仪检测SMMC-7721细胞周期的变化;Annexin V-FITC/PI双染法检测总黄酮对SMMC-7721细胞凋亡的影响。结果:MTT实验结果表明,镰形棘豆总黄酮抑制SMMC-7721细胞的增殖,并呈现出一定的浓度依赖性。Hoechst33258染色结果显示,经镰形棘豆总黄酮处理的细胞出现了多种凋亡特征和超微结构的变化。镰形棘豆总黄酮作用SMMC-7721细胞后可观察到G1期细胞明显增多,凋亡细胞的比例升高,并且呈浓度依赖性。结论:镰形棘豆总黄酮抑制SMMC-7721细胞的增殖,与其诱导细胞周期阻滞和使细胞形态发生变化而引起细胞凋亡的活性有关。值得进一步深入研究和阐述镰形棘豆抗肿瘤的作用机制。

Objective： To examine apoptosis of SMMC-7721 hepatocarcinoma cells induced by total flavonoids of Oxytropis falcata（TFOF） and its preliminary mechanism.Method： SMMC-7721 cells were treated for 24 h with TFOF in different concentrations.Inhibition on proliferation of SMMC-7721 cells was assessed by MTT assay.The morphology of treated SMMC-7721 cells was observed by optical microscope.Effect of TFOF on the nuclear morphology of cells was analyzed using Hoechst 33258 staining by fluorescence microscope.Annexin V-FITC/ PI staining and flow cytometric measurement were used for investigating the effect of TFOF on induction of apoptosis in SMMC-7721 cells and cell cycle analysis.Result： The results of MTT assay showed that TFOF could induce cytotoxicity in SMMC-7721 cells in a dose-dependent manner.Hoechst 33258 staining analysis indicated that TFOF caused typical characteristics of apoptotic programmed cell death,such as cell shrinkage,apoptotic body formation etc.Flow cytometric analysis demonstrated that TFOF caused a dose-dependent apoptosis of SMMC-7721 cells and arrested cell cycle in G1 phase.Conclusion： It suggested that TFOF inhibit proliferation of SMMC-7721 cells by inducing apoptosis of the cells and arresting cell cycle in G1 phase.