芹菜素对人肺癌NCI-H1299细胞的生长抑制和凋亡诱导的作用

The growth inhibition and apoptosis-inducing effects of apigenin on human lung cancer NCI-H1299 cells

目的研究芹菜素对人肺癌NCI-H1299细胞的生长抑制和诱导凋亡的作用。方法体外常规培养NCI-H1299细胞,芹菜素作用终浓度分别为10、20、40、80μmol/L。MTT法测定芹菜素对细胞的生长抑制作用;Hoechst 33258细胞核染色法观察细胞凋亡的形态学变化;流式细胞仪AnnexinV-FITC/PI双染色法检测细胞凋亡率;Western blot方法检测芹菜素对Bax和Bcl-2蛋白表达的影响。结果芹菜素能够抑制NCI-H1299细胞的增殖,并随药物浓度的增加、作用时间的延长,作用更明显;Hoechst 33258染色结果显示,芹菜素处理组细胞出现核固缩、染色质凝集和凋亡小体产生等典型的细胞凋亡特征;不同浓度的芹菜素作用24 h,细胞凋亡率逐渐增加,呈一定浓度依赖性;Western blot结果显示,芹菜素处理组细胞Bax蛋白表达增加,Bcl-2蛋白表达降低。结论芹菜素能够抑制人肺癌NCI-H1299细胞的增殖,并诱导其凋亡,其机制可能与调控Bax和Bcl-2的表达,导致Bcl-2/Bax比值下降有关。

Objective To investigate the growth inhibition and apoptosis-inducing effects of apigenin on human lung cancer NCI-H1299 cells.Methods The lung cancer cells NCI-H1299 were incubated with apigenin at various concentrations（10,20,40 and 80 μmol / L）.The inhibition of proliferation was assessed by MTT assay;while apoptosis of NCI-H1299 cells was observed in a fluorescence microscope by Hoechst 33258 stain,and quantified by flow cytometry using annexin V-FITC / PI stain.The expressions of apoptosis-related protein Bax and Bcl-2 were analyzed by Western blot.Results There was inhibition effects of proliferation by apigenin in concentration-and time-dependent manners.The NCI-H1299 cells treated with apigenin showed significant morphological changes in different apoptotic phases.The apigenin-induced apoptosis was concentration-dependent.Western blot analysis indicated that apigenin up-regulation of Bax,and down-regulation of Bcl-2 were induced by apigenin.Conclusion Apigenin can inhibit cell proliferation and induce apoptosis of human lung cancer cell line NCI-H1299.The mechanism may involve the up-regulation of Bax protein and down-regulation of Bcl-2 protein.