miRNA-513a通过负性调控LGR6抑制胃癌细胞侵袭和转移  

miR-513a inhibited invasion and migration of gastric cancer cells through target LGR6

作　　者：柯靖[1] 杨娟娟 章文毅[1] 毛勤生[1] 马彭[1] KE Jin;YANG Juanjuan;ZHANG Wenyi;MAO Qinsheng;MA Peng(Department of General Surgery,the Affiliated Hospital of Nantong University,Jiangsu 226001)

机构地区：[1]南通大学附属医院普外科

出　　处：《交通医学》2019年第4期348-353,共6页Medical Journal of Communications

基　　金：南通市科技项目(MS22015092)

摘　　要：目的:观察miR-513a对胃癌细胞体外侵袭-转移能力的影响,探讨miR-513a通过LGR6调节胃癌侵袭和转移的具体机制。方法:通过RT-qPCR方法检测胃癌组织及癌旁组织中miR-513a的表达;采用生物信息学预测miR-513a靶向调控的基因,筛选出LGR6。用Western blotting法检测胃癌组织及细胞中LGR6的表达变化,采用双荧光素酶实验分析miR-513a与LGR6的作用机制。采用Transwell侵袭实验及划痕实验检测miR-513a及LGR6干扰质粒转染后细胞体外侵袭能力及迁移能力的变化。结果:实时荧光定量PCR实验结果显示,miR-513a在胃癌组织中低表达,而LGR6在胃癌组织中高表达。双荧光素酶实验证实miR-513a可以结合在LGR6的3’-UTR区,miR-513a过表达后LGR6表达降低。同时miR-513a mimics转染后细胞增殖能力、侵袭能力、迁移能力均显著降低。结论:miR-513a可以在转录后水平调控LGR6的表达,从而抑制胃癌细胞的侵袭和转移。Objective: To investigate the underlying molecular mechanisms of miR-513a regulating the invasion and migration of gastric cancer cells. Methods: Expression of miR-513a in gastric cancer tissues and adjacent precancer tissues was analyzed by RT-qPCR methods. Bioinformatics was employed to predict the miRNAs related to LGR6 regulation.Western blotting was used to detect the changes of LGR6 in gastric cancer cells.Dual luciferase assay was adopted to analyze the mechanism of miR-513a regulating LGR6. Transwell invasion assay was used to analyze the effect of miR-513a on cell invasion. The quantitative real-time polymerase chain reaction and western blotting analysis were adopted to detect the expression levels of LGR6 in cells transfected with miR-NC, miR-513a mimics or miR-513a inhibitor. Results: miR-513a was in a low expression in gastric carcinoma compared with adjacent normal tissues. And bioinformatics analysis demonstrated that LGR6 was the target gene of miR-513a. Dual luciferase assay confirmed that miR-513a was bond to the 3'-UTR of LGR6. Western blotting and RT-qPCR experiments both suggested that LGR6 was down-regulated as miR-513a overexpressed. Conclusion: miR-513a regulates LGR6 expression at post-transcriptional level,and then suppresses the invasion and migration of gastric adenocarcinoma.

关 键 词：miR-513a LGR6 胃癌 肿瘤转移 

分 类 号：R573.9[医药卫生—消化系统]