芹菜素对人肺癌A549细胞凋亡及相关蛋白Bax、Bcl-2表达的影响

Effect of apigenin on apoptosis and expression of Bax and Bcl-2 in A549 human lung cancer cells

目的研究芹菜素对人肺癌A549细胞增殖抑制和凋亡诱导作用与Bax、Bcl-2蛋白表达的影响。方法 10～80μmol/L不同浓度芹菜素作用A549细胞,采用MTT法检测芹菜素对A549细胞增殖抑制作用;Hoechst 33258细胞核染色法观察芹菜素诱导细胞凋亡形态学的变化;流式细胞仪AnnexinV-FITC/PI双染色法检测细胞凋亡率;Westernblot法检测凋亡相关蛋白Bax和Bcl-2表达的变化。结果 MTT法显示,芹菜素对A549细胞有显著的增殖抑制作用(P<0.01),且具浓度和时间依赖性;荧光显微镜下观察到芹菜素处理组细胞出现典型的凋亡形态学改变:细胞核固缩、染色质凝集和核碎片化等;流式细胞仪分析结果显示,芹菜素呈浓度依赖性诱导A549细胞凋亡;Westernblot结果显示,促凋亡蛋白Bax随着芹菜素浓度升高表达增加,抗凋亡蛋白Bcl-2随着芹菜素浓度升高表达减少。结论芹菜素具有抑制人肺癌A549细胞增殖,诱导其凋亡的作用,其机制可能与上调Bax蛋白表达和下调Bcl-2蛋白表达有关。

Objective To investigate the effect of apigenin on proliferation, and apoptosis, as well as Bax and Bcl-2 protein expression in human lung cancer cell line A549. Methods A549 cells cultured in vitro were treated with 10 ~ 80 μmol/L different concentration of apigenin. MTT was used to detect the inhibitory rates according to the formula. Apoptosis of A549 cells was observed in a fluorescence microscope by Hoechst 33258 staining and quantified by flow cytometry using annexin V-FITC/PI staining. The expressions of apoptosis-related protein Bax and Bcl-2 were analyzed by Western blot. Results The inhibition of proliferation by apigenin was concentration-dependent and time-dependent. A549 cells treated with apigenin showed significant morphological changes in different apoptotic phases. The apoptotic rate was concentration-dependently increased. Western blot analysis indicated that apigenin increased the protein level of Bax, and reduced the protein level of Bcl-2. Conclusion Apigenin can inhibit proliferation and induce apoptosis of human lung cancer cell line A549, which might be related with up-regulated expression of Bax and down-regulated expression of Bcl-2.