αvβ6-ERK直接通路参与去甲斑蝥素诱导HT-29结肠癌细胞凋亡  

αvβ6-ERK pathway participates in norcantharidin-induced apoptosis of HT-29 colon cancer cells

作　　者：张琦[1] 胡燕燕[2] 彭程[3] 牛卫博[3] 刘恩宇[3] 贺兆斌[3] 赵传宗[3] 牛军[3] 

机构地区：[1]山东大学第二医院血液肿瘤科,山东济南250033 [2]山东大学齐鲁医院干部保健科,山东济南250012 [3]山东大学齐鲁医院肝胆外科,山东济南250012

出　　处：《中国病理生理杂志》2011年第8期1525-1530,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30570833;No.30872460)

摘　　要：目的:研究去甲斑蝥素(NCTD)诱导结肠癌细胞凋亡的药理学机制。方法:Hoechst 33258荧光染色检测NCTD处理后结肠癌细胞凋亡程度;流式细胞仪检测NCTD对结肠癌细胞表面整合素表达的影响;MTT检测多种功能性单抗对NCTD处理后HT-29细胞生长的影响;Western blotting分析NCTD对HT-29细胞内丝裂原活化蛋白激酶(MAPKs)表达量及磷酸化程度的影响;免疫共沉淀检测NCTD对αvβ6-ERK直接连接的影响。结果:NCTD能够诱导HT-29细胞凋亡;流式细胞术分析表明NCTD能抑制结肠癌细胞表面整合素αvβ6的表达,但对αvβ3和αvβ5的表达无明显影响;MTT实验证实αvβ6功能性单抗10D5能够增强NCTD抑制HT-29细胞生长的效应,整合素αvβ3和αvβ5的单克隆抗体LM609和F1P6无明显抑制细胞生长的能力;Western blotting分析证实,HT-29细胞经NCTD处理后仅有磷酸化ERK含量随NCTD浓度增加或作用时间延长而逐渐降低;免疫共沉淀证实NCTD干扰αvβ6-ERK直接连接的形成。结论:NCTD通过降低HT-29细胞表达整合素αvβ6,阻碍了ERK的磷酸化,干扰αvβ6-ERK直接连接形成,阻断了αvβ6介导的恶性生物学信号转导。NCTD通过αvβ6-ERK信号通路诱导人结肠癌HT-29细胞凋亡。AIM： To investigate the pharmacological mechanism of noreantharidin （.NCTD） -induced apoptosis of FIT -29 colon cancer cells. METHODS： Hoechst 33258 staining was used to analyze the apoptosis of FIT -29 cells treated with NCTI）. The effects of NCTD on the expression of integrin in HT - 29 cells were determined by flow cytometry. The effects of several functional blocking antibodies on HT - 29 cells were detected by MTT method. The expression and the phosphorylation of mitogen activated protein kinases （MAPKs） in HT - 29 ceils were measured by Western blotting. Co - immunoprecipitation assay was used to detect the activity of αvβ6 - extracellular signal - regulated kinase （ERK） direct linkage in HT - 29 cells. RESULTS ： NCTD induced the apoptosis of HT - 29 colon cancer cells. The expression of integrin αvβ6 in HT-29 cells treated with NCTD was reduced, but the expression of αvβ3 and αvβ5 was not changed. A function -blocking antibody to αvβ6,10D5, strengthened the growth inhibitory effect of NCTD on HT- 29 cells, but LM609 （ a function - blocking antibody to αvβ3 ） and P1 F6 （ a function - blocking antibody to αvβ5） did not. The level of phosphorylated ERK （ p - ERK） was decreased substantially after treated with NCTD in a dose - and time - dependent manher. NCTD also affected the association of αvβ6 and ERK. CONCLUSION： NCTD decreases the expression of integrin αvβ6 and interferes with the phosphorylation of ERK. As a result, the formation of αvβ6 - ERK direct linkage is affected and the signal transduction mediated by αvβ6 is disturbed. The mechanism of NCTD - induced HT - 29 cell apoptosis is involved in the αvβ6 -ERK signaling pathway.

关 键 词：去甲斑蝥素 整合素ΑVΒ6 细胞凋亡 结肠肿瘤 

分 类 号：R363[医药卫生—病理学]