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  

Research of Apoptosis Mechanism of NCIH460 Cells Induced by the Effective Component Quercetin in Zhoushi Kejinyan Fang

作　　者：黄震洲[1] 陈美娟[1] 熊飞[1] 姜淼[1] 尹莲[2] 张旭[1,2] 

机构地区：[1]南京中医药大学基础医学院,江苏南京210046 [2]江苏省方剂研究重点实验室,江苏南京210046

出　　处：《南京中医药大学学报》2012年第3期235-237,共3页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：科技部十一五支撑计划(2006BAI11B08-01);江苏省高校优势学科建设工程资助项目(PAPD)资助;江苏省"六大人才高峰"项目资助(2011-D-078)

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24h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25μmol/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/ERK1/2 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 NCI-H460 and inhibit the EGFR-Ras-Raf-MEK/ERK1/2 pathway by inhibiting the activation of c-Raf.

关 键 词：周氏克金岩方 槲皮素 NCI-H460细胞 ras通路 凋亡 

分 类 号：R285[医药卫生—中药学]