%0 Journal Article
%T 食管癌中MicroRNA-149与DNA聚合酶β的靶向关系<br>Target relationship between microRNA-149 and DNA polymerase β in esophageal carcinoma
%A 张
%A 满
%A 李
%A 敏
%A 赵国强
%J 郑州大学学报（医学版）
%D 2018
%R 10.13705/j.issn.1671-6825.2018.01.024
%X 目的:探讨食管癌中miR-149与DNA聚合酶β(polβ)的靶向关系。方法:采用实时荧光定量PCR方法,分析64对食管癌及其癌旁正常组织中polβ mRNA和miR-149的表达水平。采用TargetScan和miRanda进行生物信息学分析,预测miR-149和polβ的结合位点。采用Western blot法检测未转染的EC9706细胞及miR-149 mimics、miR-149 scramble转染细胞中polβ蛋白的表达。构建野生型polβ 3'UTR表达质粒pmirGLO-Wt-polβ 3'UTR, 检测miR-149 mimic或miR-149 scramble与重组质粒共转染的EC9706细胞中的荧光素酶活性。结果:与癌旁正常组织比较,食管癌组织中polβ mRNA表达水平升高, miR-149表达水平降低,两者表达呈负相关(r=-0.671,P<0.05)。miR-149 mimics转染组细胞中polβ蛋白表达水平低于miR-149 scramble转染组和未转染组(P<0.05)。miR-149 mimic与重组质粒共转染组细胞中的荧光素酶活性低于miR-149 scramble与重组质粒共转染组(P<0.05)。结论:食管癌中polβ是miR-149的靶基因,miR-149可通过结合polβ 3'UTR下调polβ的表达水平。<br>Aim: To explore the target relation between microRNA(miR)-149 and DNA polymerase β(po1β)in esophageal carcinoma.Methods: Quantitative real-time PCR was used to detect the expressions of polβ mRNA and miR-149 in 64 cases of esophageal cancer and the adjacent normal tissue. TargetScan and miRanda were used to predict the binding sites between miR-149 and polβ. Western blot was used to detect polβ protein in EC9706 cells without transfection, transfected with miR-149 mimics or miR-149 scramble. The expressing vector of wide type polβ 3'UTR was constructed, and the luciferase activity of the EC9706 cells co-transfected with the expressing vector and miR-149 mimic, or the expressing vector and miR-149 scramble.Results: Compared with those of the adjacent normal tissue, the polβ mRNA expression in tumor tissue was significantly higher, miR-149 expression was lower, and the polβ mRNA expression was negatively correlated with miR-149 expression(r=-0.671,P<0.05). The polβ protein expression in the cells transfected with miR-149 mimics was lower than the other two groups(P<0.05). The luciferase activity of the cells co-transfected with the expressing vector and miR-149 mimic was lower than those co-transfected with the expressing vector and miR-149 scramble(P<0.05).Conclusion: Polβ is a target gene of miR-149 in esophageal cancer cells, and miR-149 could negatively regulate polβ expression by binding to its 3'UTR
%K 食管癌
%K microRNA-149
%K DNA聚合酶β&lt
%K br&gt
%K esophageal carcinoma
%K microRNA-149
%K DNA polymerase β
%U http://jms.zzu.edu.cn/oa/darticle.aspx?type=view&id=201804001