Abstract
Down‐regulated in adenoma (DRA) is a Cl−/HCO3 − exchanger involved in electroneutral NaCl transport in the mammalian intestine. Altered DRA expression levels are associated with infectious and inflammatory diarrheal diseases. Therefore, it is critical to understand the regulation of DRA expression. MicroRNAs are endogenous, small RNAs that regulate protein expression via blocking the translation and/or promoting mRNA degradation. In order to investigate potential modulation of DRA expression by microRNA, five different in silico algorithms were used to predict the microRNAs that target DRA. Of these microRNAs, miR‐494 was shown to have highly conserved putative binding site in the DRA 3′untranslated region (3′UTR) compared to other DRA‐targeting microRNAs in vertebrates. Transfection with pMIR‐GLO dual luciferase vector containing DRA 3′UTR resulted in a significant decrease in relative luciferase acitivity (44.13 ± 3.7% p<0.05) as compared to empty vector. Co‐transfection of the DRA 3′UTR luciferase vector with a miR‐494 mimic resulted in markedly decreased luciferase activity (69.58 ± 2.1% p<0.05), as compared to cells transfected with negative control. The transfection of a miR‐494 mimic into Caco2 cells significantly increased the expression of miR‐494 and concomitantly decreased the DRA protein expression by (50.5 ± 5.2% p<0.05). These data demonstrate a novel regulatory pathway for DRA and indicate that targeting miR‐494 using anti‐miR‐494 could be an effective therapeutic tool for diarrheal conditions. (Supported by NIDDK/VA)
Published Version
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