黄芪对喉癌Hep-2细胞增殖和凋亡的影响

Effects of astragalus on cell proliferation and apoptosis in Hep-2 cells in vitro

关键词:  黄芪  喉癌  凋亡

Key words:  astragalus  laryngeal carcinoma  apoptosis

摘要:

[目的] 体外观察黄芪对人喉鳞状细胞癌细胞系Hep-2的抑制增殖和诱导凋亡作用并探讨其作用机制。[方法]用20、100、200 μg/mL的黄芪作用于Hep-2细胞24 h, 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,共聚焦荧光显微镜观察细胞凋亡，Western Blot检测Bcl-2和Cyclin B1的蛋白表达。[结果]黄芪对Hep-2细胞的增殖抑制作用具有明显的剂量依赖性；随着黄芪浓度增高,处于G2/M期细胞的比例逐渐升高,同时伴有G0/G1期细胞减少,细胞凋亡率逐渐升高,各实验组之间及其与对照组之间的差异均有显著性意义(P<0.05)；荧光显微镜观察可见典型细胞凋亡；Western Blot检测显示黄芪可剂量依赖性抑制Bcl-2和Cyclin B1的蛋白表达。[结论]黄芪可通过抑制Bcl-2和Cyclin B1的蛋白表达，引起喉癌细胞G2/M期阻滞，抑制增殖和凋亡,发挥抗癌作用。

Abstract:

[Objective] To investigate the mechanism underlying the anticancer activity of Astragalus on human laryngeal cancer. [Method] Hep-2 cells were treated with astragalus in different concentrations for 24 h. MTT assay was used to evaluate cell proliferation. Flow cytometry with PI staining and fluorescent microscopy with Hoechst 33258 staining were used to estimate cell cycle distribution and apoptosis. Expressions of Bcl-2 and Cyclin B1 were evaluated by western blot. [Result] Astragalus inhibited cellular proliferation in a dose dependent manner(P<0.05). Flowcytometry analysis showed that treatment with astragalus resulted in accumulation cells at the G2/ M phase of the cell cycle and cell apoptosis in a dose dependent manner (P<0.05). Marked morphological changes of cell apoptosis including condensation of chromatin, nuclear fragmentation and apoptotic bodies were observed clearly by Hoechst 33258 staining. Western blot analysis demonstrated that the expressions of Bcl-2 and Cyclin B1 were suppressed significantly. [Conclusion] Astragalus inhibited Hep-2 cells proliferation and induced them apoptosis by down regulating the expressions of Bcl-2 and Cyclin B1.