硝苯地平促进肝癌细胞Huh-7自噬小体形成并上调Beclin1的表达  

Nifedipine promotes formation of autophagosomes and enhances expression of Beclin1 in hepatoma cell line Huh-7

作　　者：周程 汪光亮 潘光玉 廖洪涛 袁树民 谭宁 ZHOU Cheng;WANG Guang-liang;PAN Guang-yu;LIAO Hong-tao;YUAN Shu-min;TAN Ning(Graduate School,Faculty of Basic Medical Sciences,Guilin Medical University,Guilin 541000,China;Dept of Histology and Embryology,Faculty of Basic Medical Sciences,Faculty of Basic Medical Sciences,Guilin Medical University,Guilin 541000,China;Dept of Biomedical Engineering,College of Biotechnology,Faculty of Basic Medical Sciences,Guilin Medical University,Guilin 541000,China;Dept of Immunology,Faculty of Basic Medical Sciences,Faculty of Basic Medical Sciences,Guilin Medical University,Guilin 541000,China;Dept of Microbiology,Faculty of Basic Medical Sciences,Guilin Medical University,Guilin 541000,China)

机构地区：[1]桂林医学院研究生院,广西桂林541000 [2]桂林医学院基础医学院组织与胚胎学教研室,广西桂林541000 [3]桂林医学院生物技术学院生物医学工程教研室,广西桂林541000 [4]桂林医学院基础医学院免疫学教研室,广西桂林541000 [5]桂林医学院基础医学院微生物学教研室,广西桂林541000

出　　处：《中国药理学通报》2022年第2期228-233,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金地区基金项目(No 81360326,81660470);广西自然科学基金面上项目(No 2019JJA140602);广西高校中青年教师科研基础能力提升项目(No 2020KY12018);广西研究生教育创新计划项目(No JGY2021138)。

摘　　要：目的探讨硝苯地平对肝癌细胞Huh-7自噬小体形成及其可能机制。方法以不同浓度的硝苯地平体外干预Huh-7细胞,通过细胞增殖实验和克隆形成实验,检测硝苯地平对Huh-7细胞增殖的影响;Western blot实验检测Huh-7细胞自噬相关蛋白Beclin1、LC3B-Ⅱ的表达变化。运用激光共聚焦显微成像技术,观测硝苯地平对Huh-7细胞自噬小体形成的作用。结果硝苯地平可明显抑制Huh-7细胞的增殖,并呈时间和浓度依赖关系,2 d时硝苯地平的IC_(50)为22.7 mg·L^(-1);浓度为25 mg·L^(-1)的硝苯地平作用使Huh-7细胞的克隆形成率较对照组明显降低,克隆形成的抑制率结果为(95.46±0.45)%;Western blot结果提示,硝苯地平明显上调Beclin1、LC3B-Ⅱ蛋白表达水平;激光共聚焦显微镜影像结果显示,硝苯地平可以促进GFP-LC3B的聚集,提示硝苯地平明显诱导Huh-7细胞自噬小体形成。结论硝苯地平明显抑制肝癌细胞Huh-7增殖,并促进Huh-7细胞自噬小体形成,其机制可能与上调Beclin1蛋白的表达有关。Aim To investigate the effect of nifedipine on the formation of autophagosomes in hepatoma cell line Huh-7 and its mechanism.Methods Different concentrations of nifedipine were used to interfere with the proliferation of Huh-7 cells in vitro.The effect of nifedipine on the proliferation of Huh-7 cells was detected by cell proliferation experiment and colony formation experiment.The expressions of Beclin1 and LC3B-Ⅱwere detected by Western blot.The effect of nifedipine on the formation of autophagosomes in Huh-7 cells was observed by laser scanning confocal microscopy.Results Nifedipine significantly inhibited the proliferation of Huh-7 cells in a time-and concentration-dependent manner.The IC_(50) of nifedipine on day 2 was 22.7 mg·L^(-1).Nifedipine at the concentration of 25 mg·L^(-1) significantly reduced the colony formation rate of Huh-7 cells compared with the control group,and the inhibition rate of colony formation was(95.46±0.45)%.Western blot analysis showed that nifedipine significantly up-regulated the protein expression levels of Beclin1 and LC3B-Ⅱ.The amount of autophagosomes in nifedipine group cells were more than that of control group,which was observed by laser scanning confocal microscopy.Conclusions Nifedipine significantly inhibits the proliferation of Huh-7 cells and promotes the formation of autophagosomes,which may be related to the up-regulation of Beclin1 protein expression by nifedipine.

关 键 词：硝苯地平 肝癌 自噬 自噬小体 BECLIN1 LC3B-Ⅱ 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R392[医药卫生—基础医学]