论文部分内容阅读
目的探讨咖啡酸苯乙酯(caffeic acid phenethyl ester,CAPE)对结肠癌HT-29细胞FAK-ERK信号转导通路中相关蛋白表达的作用,寻找其作用靶点,试图阐明CAPE抗肿瘤作用的分子机制。方法用不同浓度CAPE处理HT-29细胞,利用Hoechst33258染色法和流式细胞术,检测细胞凋亡的发生。应用Western印迹法分析不同浓度CAPE对HT-29细胞中黏着斑激酶(focal adhesion kinase,FAK)和细胞外信号调节激酶(extracellular signal-regulatedkinase,ERK)蛋白表达的影响。结果 Hoechst33258染色发现CAPE作用后凋亡细胞数量增加。流式细胞仪细胞凋亡率分析显示,0,2.55,.0,7.5和10μg/ml处理HT-29细胞24 h后,细胞凋亡率上升,呈剂量依赖性。Western印迹结果显示,在0～10μg/ml范围内不同浓度CAPE作用于HT-29细胞24 h后,FAK、ERK蛋白表达随CAPE浓度的增加而下调。结论 CAPE可诱导人结肠癌HT-29细胞凋亡,其作用机制可能与CAPE抑制FAK-ERK信号转导通路的激活有关。
Objective To investigate the effect of caffeic acid phenethyl ester (CAPE) on the expression of related proteins in the FAK-ERK signal transduction pathway of colon cancer HT-29 cells and to find out its target of action in order to elucidate the antitumor effect of CAPE mechanism. Methods HT-29 cells were treated with different concentrations of CAPE. Hoechst33258 staining and flow cytometry were used to detect the apoptosis of HT-29 cells. The effects of different concentrations of CAPE on the expression of focal adhesion kinase (FAK) and extracellular signal-regulated kinase (ERK) in HT-29 cells were analyzed by Western blotting. Results Hoechst33258 staining showed that the number of apoptotic cells increased after CAPE. Flow cytometry analysis showed that apoptosis rates of HT-29 cells treated with 0,2.55,0.0,7.5 and 10μg / ml for 24 h increased in a dose-dependent manner. Western blotting showed that the expression of FAK and ERK protein were down-regulated with the increase of CAPE concentration at different concentrations of CAPE in the range of 0 ~ 10μg / ml for 24 h. Conclusion CAPE can induce the apoptosis of human colon cancer HT-29 cells. The mechanism may be related to the inhibition of FAK-ERK signal transduction pathway by CAPE.