周氏克金岩方有效成分槲皮素促进肺癌细胞NCI -H460凋亡的相关机制研究

摘要

目的 观察周氏克金岩方有效成分槲皮素诱导肺癌细胞NCI-H460凋亡的相关作用机制.方法 采用Hoechst33258染色及透射电镜(TEM)观察槲皮素诱导细胞凋亡的形态学变化,结合Western blot法检测槲皮素对细胞内EGFR-Ras-Raf-MEK/ERK1/2及凋亡相关通路的影响.结果 各浓度的槲皮素处理细胞24 h后,以25、50μmol/L 2个药物浓度处理组的凋亡特征更为显著,荧光显微镜下可见细胞核或细胞质内浓染致密的颗粒块状荧光；在25 μmol/L槲皮素处理细胞48h后在透射电镜的观察图片中也呈现出了细胞凋亡的特征;Western blot法对信号通路相关蛋白的检测结果表明,12.5μmol/L的槲皮素能显著抑制c-Raf的活化,而在25μmol/L下能促进caspase-3和caspase-7的活化.结论 槲皮素能通过抑制c-Raf的激活,抑制EGFR-Ras-Raf-MEK/ERK1/2信号通路,同时诱导肺癌细胞凋亡.%OBJECTIVE To investigate the apoptosis mechanism of NCIH460 cells induced by quercetin, the effective component in Zhoushi Kejinyan Fang. METHODS The cellular morphology was observed by nuclear staining using Hoechst33258 and transmission electron microscopy (TEM). EGFR-Ras-Raf-MEK/ERKl/2 pathway and the apoptosis associated pathway were detected by western blotting assay. RESULTS The features of apoptosis appeared after 24 h treatment, 25 μmol/L and 50 μmol/'L quercetin were of significant apoptosis. The inhibition of c-Raf was at 12.5 μmol/L and the activation of Caspase-3 and Caspase-7 were at 25 μmol/L. CONCLUSION Quercetin could induce apoptosis of NC1-H460 and inhibit the EGFR-Ras-Raf-MEK/ERK1/2 pathway by inhibiting the activation of c-Raf.