miR-34a抑制自噬增加肝癌耐药细胞HepG2/ADM化疗敏感性  

MiR-34a inhibits autophagy and increases chemosensitivity of HepG2/ADM cells

作　　者：陈有科 高良辉 符誉[1] 黄小龙[1] CHEN You-ke;GAO Liang-hui;FU Yu;HUANG Xiao-long(Department of Hepatobiliary and Pancreatic Surgery,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院肝胆胰外科,海南海口570102

出　　处：《中国现代普通外科进展》2020年第11期841-844,共4页Chinese Journal of Current Advances in General Surgery

基　　金：2017年海南省自然科学基金项目(817331)。

摘　　要：目的:观察微小RNA-34a(mi R-34a)对肝癌耐药细胞Hep G2/ADM化疗敏感性的影响。方法:实验分为Hep G2组、Hep G2/ADM组、negative control-Hep G2/ADM组和mi R-34a mimics-Hep G2/ADM组4组,Hep G2组和Hep G2/ADM组不做特殊处理,mi R-34a mimics Hep G2/ADM组转染mi R-34a mimics,negative control-Hep G2/ADM组转染negative control RNA。荧光定量PCR检测mi R-34a基因表达。透射电子显微镜检测自噬体,Western blot检测自噬蛋白表达。CCK-8检测细胞增殖。结果:Hep G2/ADM组高倍视野下自噬体为(7.56±1.34)个,显著高于Hep G2组的(1.02±0.21)个和mi R-34a mimics-Hep G2/ADM组的(3.76±0.53)个,差异均有统计学意义(P<0.01);Hep G2/ADM组与negative control-Hep G2/ADM组间自噬体差异无统计学意义(P>0.05)。Hep G2/ADM组LC3-II/I比值为22.46±5.02,显著高于Hep G2组的2.17±0.34(P<0.01)和mi R-34a mimics-Hep G2/ADM组的7.52±1.52,差异均有统计学意义(P<0.01),Hep G2/ADM组与negative control-Hep G2/ADM组间LC3-II/I比值差异无统计学意义(P>0.05)。多柔比星对Hep G2/ADM组细胞IC50值为(23.17±4.53)μmol/L,显著高于Hep G2组的(3.02±0.42)μmol/L和mi R-34a mimics-Hep G2/ADM组的(6.13±1.05)μmol/L,差异均有统计学意义(P<0.01);多柔比星对Hep G2/ADM组与negative control-Hep G2/ADM组细胞IC50值差异无统计学意义(P>0.05)。结论:mi R-34a可以抑制Hep G2/ADM细胞自噬的激活,增加Hep G2/ADM细胞对化疗药物的敏感性。Objective: To observe the effect of mi R-34 a on chemosensitivity of HepG2/ADM cell. M ethods: The experiment was divided into: HepG2, HepG2/ADM, negative control-HepG2/ADM, and mi R-34 a mimics-HepG2/ADM group. mi R-34 a gene expression was detected by Real-time PCR. The autophagy was detected by transmission electron microscope. The autophagy protein was detected by Western blot. CCK-8 assay was used to detect growth inhibition.Results: The autophagy number in HepG2/ADM group was 7.56±1.34, which was higher than that of 1.02±0.21 in HepG2 group(P<0.01) and 3.76±0.53 in mi R-34 a mimics-HepG2/ADM group(P<0.01). There was no significant difference of autophagy number between HepG2/ADM and negative control-HepG2/ADM group(P>0.05). The ratio of LC3-II/I in HepG2/ADM group was 22.46±5.02, which was higher than that of 2.17±0.34 in HepG2 group(P<0.01) and 7.52±1.52 in mi R-34 a mimics-HepG2/ADM group(P<0.01). There was no significant difference of ratio of LC3-II/I between HepG2/ADM and negative control-HepG2/ADM group(P>0.05). The IC50 values of doxorubicin to HepG2/ADM cells was(23.17±4.53) μmol/L, which was higher than that of(3.02±0.42)μmol/L to HepG2 cells(P<0.01) and(6.13±1.05) μmol/L to mi R-34 a mimics-HepG2/ADM cells(P<0.01). There was no significant difference of ratio of IC50 values of doxorubicin to HepG2/ADM and negative control-HepG2/ADM cells(P>0.05). Conclusion: mi R-34 a can inhibit the autophagy of HepG2/ADM cells and increase the chemosensitivity sensitivity.

关 键 词：微小RNA-34a 肝癌 多药耐药 自噬 

分 类 号：R735.7[医药卫生—肿瘤]