Journal
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
Volume 55, Issue 7, Pages 512-521Publisher
SPRINGER
DOI: 10.1007/s11626-019-00366-z
Keywords
miR-194; Hnf1 beta; Proliferation; Migration; Kidney
Categories
Funding
- National Science Foundation for Young Scientists of China [31701218]
- Basic Science and Frontier Technology Research Program of Chongqing Science and Technology Commission [cstc2017jcyjA0390]
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Hepatocyte nuclear factor-1 beta (Hnf1 beta) is associated with early embryogenesis failure, renal cysts, and/or diabetes. However, factors regulating Hnf1 beta expression in metanephric mesenchyme cells remain poorly understood. Here, we analyzed the modulation relationship of Hnf1 beta and miR-194 in mouse metanephric mesenchyme (MM) cells. Bioinformatics analysis, luciferase assay and semi-quantitative real-time (qPCR), western blotting, 5-ethynyl-2 '-deoxyuridine cell proliferation assay, wound healing assay, and flow cytometry were employed to detect the function of miR-194 by targeting on Hnf1 beta in mouse MM cells. Bioinformatic prediction revealed one conserved binding site (CAGTATT) of miR-194 on Hnf1 beta 3'-UTR and luciferase reporter assay suggested that this is an effective target site of miR-194, and mutating CAGTATT with CGTACTT had no effects on luciferase activity compared with control. Overexpression of miR-194 decreased Hnf1 beta mRNA and protein level in mouse MM cells. In addition, miR-194-decreased cell proliferation and miR-194-promoted cell apoptosis and migration were reversed by overexpression of Hnf1 beta coding region. In addition, Hnf1 beta-upregulated genes were decreased in miR-194 overexpression cells and rescued in miR-194 and Hnf1 beta CDS region co-overexpression cells. Our findings explored one new regulator of Hnf1 beta and revealed the function of their regulation in cell proliferation, migration, and apoptosis in mouse metanephric mesenchyme cells. For strict regulation of Hnf1 beta in kidney development, these findings provide theoretical guidance for kidney development study and kidney disease therapy.
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