MiR-20b directly targets 3’-UTR to regulate the expression of HIF-1α

Wang Helong; Luo Yulan; Tao Xiangnan; Wang Yimeng; He Jing; Guo Shujun; Song Chuanwang

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MiR-20b directly targets 3’-UTR to regulate the expression of HIF-1α

更新时间：2023-12-22

- MiR-20b directly targets 3’-UTR to regulate the expression of HIF-1α
- Journal of Chongqing Medical University Vol. 43, Issue 5, Pages: 657(2018)
- 作者机构：
  
  1. 蚌埠医学院免疫学教研室、安徽省感染与免疫重点实验室,蚌埠,233030
  2. 蚌埠医学院第二附属医院检验科,蚌埠,233030
- 作者简介：
- 基金信息：
- DOI：
  CLC：
- Published：2018，
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Wang Helong, Luo Yulan, Tao Xiangnan, et al. MiR-20b directly targets 3’-UTR to regulate the expression of HIF-1α. [J]. Journal of Chongqing Medical University 43(5):657(2018)
DOI：

Wang Helong, Luo Yulan, Tao Xiangnan, et al. MiR-20b directly targets 3’-UTR to regulate the expression of HIF-1α. [J]. Journal of Chongqing Medical University 43(5):657(2018) DOI：

摘要

目的：探讨微小RNA-20b（microRNA-20b/miR-20b）是否直接靶向低氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）信使RNA的3’-非编码区（3’-untranslated region，3’-UTR）调控其基因表达。方法：采用TargetScan和miRanda算法预测HIF-1α 3’-UTR区是否存在miR-20b的配对序列；构建并鉴定表达HIF-1α 3’-UTR的双荧光素酶基因报告系统（pmiR-RB-ReportTM-HIF-1α 3’-UTR），将重组质粒（recombinant plasmid，RP）与mimics-miR-20b（M）或NC-miR-20b（N）共转染HeLa细胞，本研究共分为RP、RP+M、RP+N、P、P+M和P+N六组，24 h后检测各组双荧光素酶活性的比值。结果：HIF-1α 3’-UTR区存在miR-20b配对区域；成功构建了含HIF-1α 3’-UTR序列的双荧光素酶基因报告载体；重组质粒与mimics-miR-20b共转染的HeLa细胞荧光素酶活性明显低于其他对照组（P=0.022）。结论：miR-20b可以直接靶向结合HIF-1α 3’-UTR区负向调控其表达。

Abstract

Objective：To investigate whether miRNA-20b（miRNA-20b/miR-20b） may bind to the 3’-untranslated region（3’-UTR） of hypoxia inducible factor-1α（HIF-1α） mRNA to regulate the expression of HIF-1α. Methods：Targetscan and miRanda softwares were used to predict the presence or absence of miR-20b binding site in the HIF-1α 3’-UTR region. The dual luciferase gene re－porter assay system（pmiR-RB-ReportTM-HIF-1α） was constructed and identified. The ratios of dual luciferase activity were detected in HeLa cells 24 hours after mimics-miR-20b（M） or NC-miR-20b（N） and recombinant plasmid（RP） co-transfection. This study was divided into six groups：RP，RP+M，RP+N，P，P+M and P+N groups. Results：There was a miR-20b binding site in the 3’-UTR of HIF-1α mRNA. The double luciferase gene reporter vector containing HIF-1α 3’-UTR was successfully constructed. The dual lu－ciferase activity was significantly lower in recombinant plasmid and mimics-miR-20b co-transfection group than in empty group（P=0.022）. Conclusion：miR-20b can directly bind to HIF-1α 3’-UTR region to regulate the expression of HIF-1α.

关键词

低氧诱导因子3’-非编码区miR-20b双荧光素酶基因报告系统

Keywords

hypoxia inducible factor3’-untranslated regionmiR-20bdual luciferase gene reporting system

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