食管癌EC9706细胞中miR-149 rs140584242多态性对DNA聚合酶β表达的影响  

Effects of miR-149 rs140584242 polymorphism on expression of DNA polymeraseβin esophageal cancer EC9706 cells

作　　者：张晓红 张文文 臧文巧 赵国强 ZHANG Xiaohong;ZHANG Wenwen;ZANG Wenqiao;ZHAO Guoqiang(Department of Basic Medicine,Zhengzhou Health Vocational College, Zhengzhou 450005;Department of Immunology,School of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450001)

机构地区：[1]郑州卫生健康职业学院基础医学系,郑州450005 [2]郑州大学基础医学院免疫学系,郑州450001

出　　处：《郑州大学学报（医学版）》2020年第3期327-330,共4页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(81672755)。

摘　　要：目的:探讨食管癌EC9706细胞中miR-149 rs140584242(G→A)多态性对调控DNA聚合酶β(polβ)表达的影响。方法:分别合成野生型miR-149和rs140584242多态性miR-149,以miR-NC为对照。采用表面等离子共振实验、双荧光素酶报告实验和Western bolt法,分析多态性miR-149与polβ3’UTR结合能力的变化及对EC9706细胞中polβ蛋白表达的影响。结果:多态性miR-149与野生型polβ的结合力低于野生型miR-149(P<0.05)。多态性miR-149与野生型polβ3’UTR报告基因载体共转染的荧光素酶活性高于野生型miR-149(P<0.05)。多态性miR-149转染的EC9706细胞中polβ蛋白相对表达量高于野生型miR-149(P<0.05)。结论:在食管癌EC9706细胞中rs140584242多态性可导致miR-149对polβ表达的负调控作用丧失。Aim:To investigate the effects of miR-149 rs140584242(G→A)polymorphism on the expression of DNA polymeraseβ(polβ)in esophageal cancer EC9706 cells.Methods:Polymorphic miR-149 rs140584242 was synthesized.EC9706 cells were transfected with wild type miR-149,miR-149 rs140584242,and miR-NC(negative control).Surfaceplasmon resonance test,dual luciferase reporter assay and Western blot were conducted to compare the binding capacity as well as the influence on polβexpression between wild type and polymorphic miR-149.Results:The binding capacity of polymorphic miR-149 to wild type polβwas significantly lower compared with wild type miR-149(P<0.05).The luciferase activity of the cells co-transfected with polymorphic miR-149 and wild type polβ3’UTR reporter gene vector was significantly higher compared with that in wild type miR-149 group(P<0.05).The expression of polβprotein in EC9706 cells transfected with polymorphic miR-149 was higher than that in wild type miR-149 group(P<0.05).Conclusion:The polymorphism of rs140584242 in esophageal cancer EC9706 cells leads to a loss of negative regulation of polβexpression by miR-149.

关 键 词：食管癌 miR-149 DNA聚合酶Β EC9706细胞 

分 类 号：R735.1[医药卫生—肿瘤]