Effect and mechanism of miR-206/miR-613 on expression of OATP1B1
10.16438/j.0513-4870.2016-0477
- VernacularTitle:miR-206/miR-613对OATP1B1基因和蛋白表达的影响及其机制研究
- Author:
Yun RAO
1
;
Gao-feng JIN
1
;
Ming-yi LIU
1
;
Xin-hua LI
1
;
Hong ZHANG
1
;
Chun-hua XIA
1
;
Yu-qing XIONG
1
Author Information
1. Clinical Pharmacology Institute of Nanchang University, Nanchang 330006, China
- Publication Type:ORIGINAL ARTICLES
- Keywords:
miR-206;
miR-613;
organic anion transporting polypeptide 1B1;
post-transcriptional regulation
- From:
Acta Pharmaceutica Sinica
2016;51(12):1858-
- CountryChina
- Language:Chinese
-
Abstract:
This study was designed to explore the effect and mechanism of miR-206/miR-613 on the expression of OATP1B1 gene. Bioinformatic analysis was used to predict the potential miRNAs target sites in 3'-untranslated region (3'-UTR) of OATP1B1 mRNA. The expression level of miR-206/miR-613 and OATP1B1 mRNA and protein was determined with RT-qPCR and Western blot, respectively. Luciferase assay was used to explore the exact mechanism of the effect of miR-206/miR-613 on the expression of OATP1B1 mRNA and protein. The results showed that the seed sequences of miR-206/miR-613 has perfect complementary with 3'-UTR of OATP1B1 mRNA in terms of sequence specificity. The secondary structure between miR-206/miR-613 and 3'-UTR of OATP1B1 mRNA was rather stable. The OATP1B1 protein level was down-regulated by 24.7%, 38.8% by overexpression of miR-206/miR-613. The expression was up-regulated by 25%, 38.2% by inhibition of miR-206/miR-613. However, overexpression or inhibition of miR-206/miR-613 had no effect on the expression of OATP1B1 mRNA. The luciferase activity of pMIR/OATP1B1-WT luciferase reporter gene was decreased by 35% and 30% through overexpression of miR-206/miR-613. The expression was increased by 33.1% and 32.5% through inhibition of miR-206/miR-613. When the binding sites in the 3'-UTR of OATP1B1 mRNA complementary with miR-206/miR-613 was mutated, overexpression or inhibition of miR-206/miR-613 had no effect on the luciferase activity. Collectively, miR-206/miR-613 post-transcriptionally regulates the expression of OATP1B1 protein by directly targeting the 3'-UTR of OATP1B1 mRNA.
