miR-143调控DNMT3B表达对肝细胞癌耐阿霉素敏感性的影响  

Effect of the DNMT3B expression on regulated by miR-143 the sensitivity of the hepatocellular carcinoma resistance to adriamycin

作　　者：崔兵[1,2] 韩正阳 万诚诚[1] 刘纪君 贺晓珊[1] 梅传忠 CUI Bing;HAN Zheng-yang;WAN Cheng-cheng;LIU Ji-jun;HE Xiao-shan;MEI Chuan-zhong(Clinical Testing and Diagnose Experiment Center,3. Department of Biochemistry and Molecular Biology ,Bengbu Medical College, Bengbu Anhui 233030;Department of Blood Transfusion, The Affiliated Hospital of Jiangsu University ,Zhenjiang Jiangsu 212001, China)

机构地区：[1]蚌埠医学院临床检验诊断学实验中心,安徽蚌埠233030 [2]江苏大学附属医院输血科,江苏镇江212001 [3]蚌埠医学院生物化学与分子生物学教研室,安徽蚌埠233030

出　　处：《蚌埠医学院学报》2018年第2期141-145,共5页Journal of Bengbu Medical College

基　　金：安徽省高校自然科学研究重点项目(KJ2016A467);安徽省教育厅自然科学研究项目(KJ2011B094);蚌埠医学院科技发展基金项目(Bykf12A01);蚌埠医学院研究生科研创新项目(Byycx1410)

摘　　要：目的:构建耐药细胞株SMMC-7721/ADM,探讨miR-143在调控肝细胞癌耐阿霉素中的作用机制。方法:采用逐步递增阿霉素浓度法构建耐阿霉素细胞株SMMC-7721/ADM,评估其IC_(50)值;采用荧光定量PCR技术检测miR-143、DNA甲基转移酶3B(DNMT3B)、多耐药基因1(MDR1)变化、Bax和Bcl-2 mRNA表达水平;Western blotting检测DNMT3B蛋白表达;通过转染miR-143 mimics上调耐药细胞株SMMC-7721/ADM中miR-143表达水平后,检测耐药细胞IC_(50)、侵袭能力、细胞凋亡、DNMT3B、MDR1变化。应用脂质体-2000瞬时转染miR-143 mimics于耐药细胞株,观察上述指标变化。结果:成功构建耐药细胞株SMMC-7721/ADM,IC_(50)值升高78.97倍(P<0.01),miR-143表达量降低,DNMT3B、MDR1、Bax、Bcl-2表达量升高(P<0.05~P<0.01)。转染miR-143 mimics后耐药细胞株IC_(50)值降低57%,侵袭能力下降,细胞凋亡增加,DNMT3B表达减少,MDR1 mRNA表达减少(P<0.01)。结论:miR-143可能通过降低DNMT3B表达提高肝细胞癌对阿霉素的敏感性。Objective： To establish hepatocellular carcinoma cell line SMMC-7721/ADM resistance to adriamycin,and explore the mechanism of microRNA（ miR）-143 in regulating the cell line resistance to adriamycin. Methods： The cell line SMMC-7721/ADM resistance to adriamycin was constructed using the gradual increasing the concentration of adriamycin,and the IC50 value of which was evaluated. The mRNA expression levels of miR-143,DNA methyltransferase 3 B（ DNMT3 B） and multidrug resistance gene 1（ MDR1）,Bax and Bcl-2 genes were evaluated using real-time fluorescent quantitative PCR,and the protein level of DNMT3 B was determined by Western blotting. After the miR-143 mimics upregulating the miR-143 expression in SMMC-7721/ADM cell line,the IC50 value,invasion ability,cell apoptosis,and levels of DNMT3 B,MDR1 were detected. The miR-143 mimics was transiently transfected into the drug resistance cell line using liposome-2000,and the indexes changes were observed. Results： The cell line SMMC-7721/ADM resistance to adriamycin was successfully established,the IC50 value raised 78. 97 times（ P 〈0. 01）. The expression level of miR-143 decreased,and the expression levels of DNMT3 B,MDR1,Bax and Bcl-2 increased,and the differences of which were statistically significant（ P 〈0. 05 to P 〈0. 01）. After the miR-143 mimics transfecting into the drug resistance cell line,the IC50 value reduced57%,the invasion ability declined,the cell apoptosis increased,the levels of DNMT3 B and MDR1 mRNA decreased,and the differences of which were statistically significant（ P〈 0. 01）. Conclusions： The increasing sensitivity of hepatocellular carcinoma to adriamycin may be caused by the miR-143 reducing DNMT3 B expression.

关 键 词：肝肿瘤 阿霉素 MIR-143 DNA甲基转移酶3B 耐药 

分 类 号：R737.9[医药卫生—肿瘤]