Identification and Functional Verification of MicroRNA-16 Family Targeting Intestinal Divalent Metal Transporter 1 (DMT1) in vitro and in vivo

• Published in: Frontiers in physiology Vol. 10; p. 819
• Main Authors: Jiang, Shuxia, Guo, Shihui, Li, Huifang, Ni, Yingdong, Ma, Wenqiang, Zhao, Ruqian
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 27.06.2019
• Subjects: DMT1; functional verification; intestine; microRNA-16 family; Physiology; regulation; functional verification; intestine; regulation; DMT1; microRNA-16 family
• ISSN: 1664-042X; 1664-042X
• DOI: 10.3389/fphys.2019.00819

Divalent metal transporter 1 (DMT1) is a key transporter of iron uptake and delivering in human and animals. However, post-transcriptional regulation of DMT1 is poorly understood. In this study, bioinformatic algorithms (TargetScan, PITA, miRanda, and miRDB) were applied to predict, screen, analyze, and obtain microRNA-16 family members (miR-16, miR-195, miR-497, and miR-15b) targeting DMT1, seed sequence and their binding sites within DMT1 3' untranslated region (3' UTR) region. As demonstrated by dual-luciferase reporter assays, luciferase activity of DMT1 3' UTR reporter was impaired/enhanced when microRNA-16 family member over-expression plasmid/its inhibitor was transfected to HCT116 cells. Corroboratively, co-transfection of microRNA-16 family member over-expression plasmid and DMT1 3' UTR mutant reporter repressed the luciferase activity in HCT116 cells. In addition, over-expression microRNA-16 family member augmented its expression and diminished DMT1 protein expression in HCT116 cells. Interestingly, tail vein injection of miR-16 assay revealed reduced plasma iron levels, higher miR-16 expression, and lower DMT1 protein expression in the duodenum of mice. Taken together, we provide evidence that microRNA-16 family (miR-16, miR-195, miR-497, and miR-15b) is confirmed to repress intestinal DMT1 expression and , which will give valuable insight into post-transcriptional regulation of DMT1.