黄芪对喉癌Hep-2细胞增殖与转移能力的影响

Effects of Astragalus on cell proliferation and metastasis in Hep-2 cell line in vitro

目的:体外观察黄芪对人喉癌细胞系Hep-2的抑制增殖和转移能力的作用并探讨其作用机制。方法:用20、100、200μg/ml的黄芪作用于Hep-2细胞24h,MTT法检测细胞增殖,流式细胞仪检测细胞周期分布和细胞凋亡率,侵袭实验观察药物对Hep-2细胞侵袭转移能力影响,AO/EB染色观察细胞凋亡,Westernblot检测Bcl-2和Bax蛋白的表达。结果:MTT结果显示黄芪对Hep-2细胞的增殖抑制作用具有明显的剂量依赖性;流式细胞仪检测发现随着黄芪浓度增高,细胞凋亡率逐渐升高,统计学分析,各实验组之间及其与对照组之间的差异均有统计学意义(P<0.05);AO/EB染色后可见典型细胞凋亡的形态变化;Hep-2细胞体外侵袭能力明显受抑制,存在剂量依赖性;Western blot检测显示黄芪可剂量依赖性地抑制Bcl-2蛋白表达。结论:黄芪可通过抑制Bcl-2蛋白表达,上调Bax表达,引起喉癌细胞增殖抑制和凋亡,发挥抗癌作用。

Objective：To investigate the mechanism underlying the anticancer activity of Astragalus on human laryngeal cancer.Methods：Hep-2 cells were treated with different concentrations of Astragalus for 24h.MTT assay was used to evaluate cell proliferation.Flow cytometry with PI staining.Invasion assay was used to measure the effect of metastasis function by using Astragalus and fluorescent microscopy with AO/EB staining were used to estimate cell cycle distribution and cell apoptosis.Expression of Bcl-2 and Bax proteins was evaluated by Western blot assay.Results：Astragalus inhibited cellular proliferation in a dose dependent manner.Flowcytometry analysis showed that treatment with Astragalus resulted in cell apoptosis in a dose dependent manner.The invasion function was inhibited significantly in a dose dependent manner.Marked morphological changes of cell apoptosis including condensation of chromatin,nuclear fragmentation and apoptotic bodies were observed clearly by AO/EB staining.Western blot analysis demonstrated that the expression of Bcl-2 proteins was suppressed significantly.Conclusion：Astragalus inhibited cell proliferation and induced apoptosis of Hep-2 cells by down regulating the expression of Bcl-2 and up regulating the expression of Bax proteins.