咖啡酸苯乙酯靶向调控人结肠癌HT-29细胞FAK-ERK信号通路的研究

Caffeic acid phenethyl ester induces apoptosis of colon cancer cells HT-29 by inhibiting FAK/ERK signal transduction pathway

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 explore the effect of caffeic acid phenethyl ester（CAPE） on the expression of related proteins in focal adhesion kinase（FAK）-extracellular signal-regulated kinase（ERK） signal transduction pathway in colon cancer cell line HT-29,to find the targets and to elucidate the anti-tumor mechanism of CAPE.Methods The cells of human colon cancer cell line HT-29 were treated with CAPE at different concentrations.Flow cytometry（FCM）and Hoechst33258 staining were used to detect apoptosis.Western-blot analysis was used to evaluate the protein level of FAK-ERK in HT-29 cells treated by CAPE.Results Hoechst33258 staining showed that apoptosis cells increased after the treatment of CAPE.The results of FCM analysis showed that the cell apoptosis rate increased after exposure to CAPE in a dose dependent manner（0,2.5,5.0,7.5 and 10 μg/ml） after 24 h.Western-blot analysis showed that the expression of FAK-ERK protein in HT-29 cells was down-regulated with the increase in CAPE concentrations from 0 to 10 μg/ml.Conclusion CAPE can induce the apoptosis of the colon cancer cell line HT-29.The mechanism of CAPE is to suppress FAK-ERK signal transduction pathway activation.