抗菌肽LL-37通过激活AMPK介导的自噬对结肠癌HT-29细胞凋亡的研究  

Study of antimicrobial peptide LL-37 on apoptosis of colon carcinomacell line HT-29 by activating AMPK-mediated autophagy

作　　者：谢亚锋 许志杰 丁雅婷 闫圣玉 张侨 刘菀莹 刘丽兵[2] Xie Yafeng;Xu Zhijie;Ding Yating(Dept of Anorectal,The Second Affiliated Hospital of Nanhua Uniersity,Hengyang421001)

机构地区：[1]南华大学附属第二医院肛肠科,衡阳421001 [2]南华大学附属南华医院肛肠科,衡阳421001

出　　处：《安徽医科大学学报》2020年第12期1845-1849,共5页Acta Universitatis Medicinalis Anhui

基　　金：湖南省教育厅科学研究项目(编号:16C1408);衡阳市科技局2018年指导性项目(编号:S2018F9031022235)。

摘　　要：目的探讨抗菌肽LL-37对结肠癌HT-29细胞凋亡的影响及其分子机制。方法以结肠癌HT-29细胞作为研究对象,加入不同浓度(50、100、200μg/ml)的抗菌肽LL-37或联合AMP依赖的蛋白激酶(AMPK)抑制剂多索吗啡(10μmol/L)处理48h后,采用流式细胞术检测HT-29细胞凋亡率,Westernblot实验检测HT-29细胞凋亡相关蛋白(Bcl-2和Bax)、自噬相关蛋白(Beclin1、Atg5、LC3I和LC3II)和AMPK蛋白的表达。结果高浓度抗菌肽LL-37能促进结肠癌HT-29细胞凋亡(P<0.05),并上调自噬相关蛋白Bec-lin1、Atg5表达水平以及提高LC3II/LC3I值(P<0.05),并且伴随着AMPK异常激活(P<0.01);AMPK抑制剂联合干预能逆转抗菌肽LL-37对HT-29细胞的作用效果。结论抗菌肽LL-37通过激活AMPK介导的自噬促进结肠癌HT-29细胞的凋亡。Objective To investigate the effect of antimicrobial peptide LL-37 on apoptosis of colon carcinoma HT-29 cells and its molecular mechanism. Methods Colon cancer HT-29 cells were treated with antimicrobial peptide LL-37 at different concentrations of 50 μg/ml,100 μg/ml and 200 μg/ml or LL-37 combined with AMPK inhibitor dorsomorphin(10 μmol/L, pre-treated for 30 min) for 48 h. The apoptosis rate of HT-29 cells was assessed by flow cytometry. Apoptotic proteins(Bcl-2 and Bax), autophagy-related proteins(Beclin1, Atg5, LC3 I and LC3 II) and AMPK protein were detected by Western blot assay. Results High dose antimicrobial peptide LL-37 could promote apoptosis of HT-29 cells in colon carcinoma(P<0.05), and increase the expression of autophagy related proteins Beclin1, Atg5 and LC3 II/LC3 I(P<0.05), accompanied by abnormal activation of AMPK(P<0.01). Intervention of AMPK inhibitor dorsomorphin combined with LL-37 could reverse the effect of antimicrobial peptide LL-37 on HT-29 cells in colon carcinoma. Conclusion Antimicrobial peptide LL-37 promotes apoptosis of colon cancer cell line HT-29 by activating AMPK-mediated autophagy.

关 键 词：结肠癌HT-29细胞 AMPK 抗菌肽LL-37 细胞自噬 细胞凋亡 

分 类 号：R735.55[医药卫生—肿瘤]