摘要
目的探讨胃癌中MIRNA-199A-3P调控候选基因MAP3K4的表达。方法收集35例胃癌及癌旁组织标本,采用茎环逆转录-聚合酶链式反应法检测胃癌组织及癌旁组织中MIR-199A-3P的表达水平,WESTERN BLOT法检测MAP3K4表达;细胞转染法检测MIR-199A-3P调控MAP3K4表达,双荧光素酶报告基因检测系统验证MIR-199A-3P结合MAP3K4的3’UTR的靶位点。结果与癌旁组织相比,胃癌组织MIR-199A-3P高表达,且其候选基因MAP3K4低表达;MIR-199A-3P MIMICS转染抑制MAP3K4表达;MAP3K4是MIR-199A-3P作用的靶基因。结论 MIR-199A-3P是胃癌的一个癌基因,MAP3K4是受其调控的靶基因。
Objective To investigate the effect of miRNA-199a-3p overexpression on the expression of MAP3K4 protein in gastric cancer. Methods 35 gastric cancers and the matched adjacent tissue specimens were collected. Expression of miRNA-199a-3p and MAP3K4 were detected by stem-loop real-time reverse transcription polymerase chain reaction and western blot. Cell transfection was employed to explore the regulation of miRNA-199a -3p on MAP3K4 gene. Luciferase reporter assay was performed to identify target MAP3K4 gene. Results Compared with the adjacent tissue specimens, miRNA-199a-3p was upregulated in the gastric cancers, and MAP3K4 protein was down-regulated in the gastric cancers. Cells transfected with miR- 199a-3p mimics showed lower MAP3K4 protein. MAP3K4 was identified as target gene of miR-199a-3p. Conclusions miRNA-199a-3p acts as an oncogene in gastric cancer and functions by targeting MAP3K4.
出处
《实用医学杂志》
CAS
北大核心
2017年第17期2817-2820,共4页
The Journal of Practical Medicine
基金
武汉市卫计委科研基金资助项目(编号:WX13C15)
