miR-200c抑制自噬对肝癌耐药细胞HepG2/ADM化疗敏感性的影响  

MiR-200c 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(Department of hepatobiliary and pancreatic surgery,the First Affiliated Hospital of Hainan Medical College,Haikou Hainan,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：《中西医结合肝病杂志》2020年第6期524-526,共3页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：海南省自然科学基金项目(No.817331)。

摘　　要：目的:观察微小RNA-200c(miR-200c)调节自噬对肝癌耐药细胞HepG2/ADM化疗敏感性的影响。方法:肝癌及肝癌耐药细胞分为4组,即HepG2组、HepG2/ADM组、阴性转染对照组和miR-200c类似物转染组,HepG2组和HepG2/ADM组不进行质粒转染,阴性转染对照组HepG2/ADM细胞转染阴性对照质粒miRNA-pEZ-M02-UBQLN1,miR-200c类似物转染组HepG2/ADM细胞转染miR-200c mimics-pEZ-M02-UBQLN1质粒。荧光定量PCR检测各组细胞miR-200c基因的表达,MTT检测各组细胞增殖情况,透射电子显微镜检测各组细胞自噬体,Western Blot检测自噬相关蛋白Beclin-1和LC3-Ⅱ的表达。结果:HepG2/ADM组细胞miR-200c相对表达量显著低于HepG2组(P<0.01),miR-200c类似物转染组miR-200c相对表达量显著高于HepG2/ADM组(P<0.01)。多柔比星对miR-200c类似物转染组细胞IC50值显著低于HepG2/ADM组(P<0.01)。HepG2/ADM组细胞高倍视野下自噬体显著多于HepG2组(P<0.01),miR-200c类似物转染组细胞自噬体显著少于HepG2/ADM组(P<0.01)。HepG2/ADM组细胞Beclin-1和LC3-Ⅱ蛋白相对表达量显著高于HepG2组(P<0.01),miR-200c类似物转染组Beclin-1和LC3-Ⅱ蛋白相对表达量显著低于HepG2/ADM组(P<0.01)。结论:上调miR-200c表达可以抑制HepG2/ADM细胞自噬,增加HepG2/ADM细胞对化疗药物的敏感性。Objective: To observe the effect of miR-200 c on chemosensitivity of HepG2/ADM cell by regulating autophagy.Methods: The experiment was divided into: HepG2, HepG2/ADM, negative control plasmid-HepG2/ADM,and miR-200 c mimics plasmid-HepG2/ADM group. The miR-200 c gene was detected by Real-time PCR.MTT assay was used to detect growth inhibition.The autophagy was detected by transmission electron microscope.The protein was detected by Western blot. Results:The 2^-△△CT values of miR-200 c in HepG2/ADM group was lower than that in HepG2 group(P<0.01).The 2^-△△CTvalues of miR-200 c in miR-200 c mimics plasmid-HepG2/ADM group was higher than that in HepG2 group(P<0.01).The IC50 values of doxorubicin to miR-200 c mimics plasmid-HepG2/ADM cells was lower than that in HepG2 group(P<0.01).The autophagy in HepG2/ADM group was higher than that in HepG2/ADM group(P<0.01). The autophagy in miR-200 c mimics plasmid-HepG2/ADM group was lower than that in HepG2/ADM group(P<0.01).The Beclin-1 and LC3-Ⅱ expression in HepG2/ADM group was higher than that in HepG2/ADM group(P<0.01).The Beclin-1 and LC3-Ⅱ expression in miR-200 c mimics plasmid-HepG2/ADM group was higher than that in HepG2/ADM group(P<0.01).Conclusion:miR-200 c can inhibit the autophagy of HepG2/ADM cells and increase the chemosensitivity sensitivity.

关 键 词：微小RNA-200c 肝细胞癌 多药耐药 自噬 

分 类 号：R735.7[医药卫生—肿瘤]