摘要
目的检测小檗碱对食管癌细胞增殖、周期和凋亡的影响,并探讨其作用机制。方法 MTT法检测细胞的增殖抑制作用;碘化丙啶(Propidium iodide,PI)染色法检测细胞周期进程;Annexin V-FITC和PI双染法结合流式细胞技术检测小檗碱的凋亡诱导作用;Western blot技术检测小檗碱对胰岛素样生长因子1受体(IGF-1R)的活化及下游主要信号分子AKT和p44/42MAPK(ERK)磷酸化水平的影响。结果小檗碱在体外可浓度依赖性地抑制四种食管癌细胞系的增殖,且其IC50值与细胞表面IGF-1R的表达水平呈负相关。小檗碱可使细胞阻滞在G2/M期。细胞凋亡实验结果显示,小檗碱可浓度依赖性地诱导食管癌细胞发生凋亡,并且在相同浓度下KYSE450细胞(IGF-1R高表达)的凋亡率显著高于KYSE150细胞(IGF-1R低表达)。Western blot结果显示,小檗碱处理可显著抑制IGF-1R的磷酸化,并抑制AKT和ERK的活化,且抑制作用随着小檗碱浓度的增加而增强。结论小檗碱可通过对IGF-1R及其介导的下游信号通路的调控来发挥抑制食管癌细胞增殖、阻断细胞周期进程以及诱导食管癌细胞凋亡的作用。
Objective To investigate the effects of berberine on the proliferation, cell cycle progression and apoptosis of esophageal cancer cells and related mechanism. Methods MTT assay was used to evaluate the effects of berberine on cell viability. Propidium iodide staining was used to detect cell cycle progression. Annexin V-FITC/PI staining assay was used to measure cell apoptosis, and Western blot analysis was performed to determine the phosphorylation of insulin-like growth factor 1 receptor(IGF-1 R) as well as the two major downstream signaling molecules AKT and p42/44 MAPK(ERK). Results In vitro, berberine inhibited the proliferation of four esophageal cancer cell lines in a concentration-dependent manner. Moreover, the IC50 values of berberine against different esophageal cancer cells were negatively correlated with the expression level of IGF-1 R. The distribution of KYSE450 cells cycle was arrested in G2/M phase. Berberine dosedependently induced the cell apoptosis, and the apoptosis ratio of KYSE450 cells with high IGF-1 R level was significantly higher than that of KYSE150 cells with low IGF-1 R level after berberine treatment at the same concentration. Berberine dose-dependently inhibited the phosphorylation of IGF-1 R and blocked AKT and ERK activation. Conclusion Berberine could inhibit cell proliferation, cell cycle arrest and induce cell apoptosis of esophageal cancer via inactivation of IGF-1 R as well as the downstream molecules AKT and ERK.
作者
李赛赛
郭晓芳
朱小飞
豆巧华
LI Saisai;GUO Xiaofang;ZHU Xiaofei;DOU Qiaohua(Department of Microbiology, School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang 453003, China;Department of Clinical Immunology, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, China)
出处
《肿瘤防治研究》
CAS
CSCD
2018年第6期362-366,共5页
Cancer Research on Prevention and Treatment
基金
河南省高校科技创新人才项目(15HASTIT040)
新乡医学院研究生科研创新支持计划(YJSCX2016082)
关键词
小檗碱
食管癌细胞
胰岛素样生长因子1受体
细胞凋亡
Berberine
Esophageal cancer cells
Insulin-like growth factor 1 receptor
Cell apoptosis