木犀草素诱导非小细胞肺癌细胞株A549凋亡和G_2周期阻滞  

Mechanism of growth inhibition effect of 3′,4′,5,7-tetrahydroxyflavone on A549 cells

作　　者：胡春萍[1] 蔡雪婷[1] 胡婷婷[2] 卢悟广[1] 曹鹏[1] 

机构地区：[1]江苏省中医药研究院细胞与分子生物学实验室,江苏南京210028 [2]南京市江北人民医院,江苏南京210048

出　　处：《中国中药杂志》2012年第9期1259-1264,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30873410);江苏省中医药领军人才项目

摘　　要：目的:研究木犀草素诱导非小细胞肺癌细胞株A549细胞凋亡和抑制其细胞周期(G2期)的分子机制。方法:MTT法分析木犀草素对A549细胞的生长抑制作用和半数抑制率IC50。Hoechst 33258核染色,Annexin V-FITC/PI双染,流式细胞术检测细胞周期和凋亡,Western blot分析木犀草素引起的周期相关蛋白和凋亡相关蛋白的变化。Western blot和免疫细胞化学推测可能存在的分子机制。结果:木犀草素对A549细胞有显著的生长抑制作用,48 h的IC50为45.2μmol.L-1,流式细胞术检测细胞周期发现A549细胞经木犀草素处理后主要阻滞在G2期,周期蛋白cyclin A,p-CDC2和p-Rb都呈低表达。Hoechst 33258核染色,Annexin V-FITC/PI双染发现木犀草素处理组细胞凋亡率明显高于未处理组。Western blot发现木犀草素可上调JNK的磷酸化,下调NF-κB(p65)的磷酸化水平。免疫细胞化学显示木犀草素可抑制TNF-α刺激的p65入核,抑制其入核发挥转录因子的作用,促进细胞凋亡。结论:木犀草素能显著诱导人非小细胞肺癌细胞A549细胞凋亡和细胞周期阻滞,其可能的分子机制是通过上调JNK磷酸化继而激活线粒体凋亡途径,同时抑制NF-κB入核使其不能发挥转录活性。Objective： To study luteolin-induced non-small cell lung cancer cell line A549 apoptosis and the molecular mechanism for inhibiting its cycle arrest（G2 stage）.Method： MTT assay showed that luteolin had obvious inhibitory effect on A549 and indicated the half inhibition ratio（IC50）.Cell cycle and apoptosis were detected by Hoechst 33258 nuclear staining assay,Annexin V-FITC/PI double staining and flow cytometry.Western blotting assay revealed changes in cycle and apoptosis-related proteins induced by luteolin.Possible molecular mechanism was suggested by Western blotting and immunocytochemistry.Result： Luteolin had an obvious growth inhibitory effect on A549 cells,with IC50 of 45.2 μmol·L-1 at 48 h.Flow cytometry showed A549 cells mainly arrested in G2 stage after being treated by luteolin,with low expressions in cyclin A,p-CDC2 and p-Rb.Hoechst 33258 nuclear staining and Annexin V-FITC/PI double staining showed that the luteolin treatment group showed a significant apoptosis rate than the non-treatment group.Western blotting found luteolin can increase phosphorylation of JNK and decrease that of NF-κB（p65）.Immunocytochemistry results revealed luteolin can inhibit TNF-α-stimulated p65 from nuclear translocation as a transcription factor and thus promoting cell apoptosis.Conclusion： luteolin can obviously induce apoptosis of human non-small cell lung cancer cell A549 possibly by increasing phosphorylation of JNK to activate mitochondria apoptosis pathway,while inhibiting NF-κB from nuclear translocation as a transcription factor.

关 键 词：非小细胞肺癌 A549 木犀草素 细胞周期 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学] R734.2[医药卫生—中医学]