MiR-193b-3p regulates chondrogenesis of ATDC5 cells via targeting TGFBR3

Abstract

Purpose: To investigate the biological effect of mmu-miR-193b-3p on chondrogenic differentiation. Methods: Chondrocyte-like ATDC5 cell line was stimulated with ITS+ premix to form cartilage nodules. Total RNA was isolated and reverse transcripted into cDNA. The 3’-UTR of predicted target genes, TGFBR3, were cloned into luciferase reporter plasmids. The mmu-miR-193b-3p mimic/inhibitor, and luciferase reporter plasmids were transfected into cells with lipofectamine 2000. Alcian blue were used to stain the cartilage nodules. Results: The miR-193b expression was elevated in chondrogenic ATDC5.The miR-193b suppressed the expression of several chondrogenic markers in chondrogenic ATDC5 in a dose dependent manner, including col2a1, sox9, col10a1, col11a1, runx2, aggrecan, and comp. The mouse TGFBR3 were predicted as the potential target gene of mmu-miR-193b-3p. The luminescence decreased more than 30% in 3T3 cells cotransfected with TGFBR3 3’-UTR reporter plasmids and miR-193b-3p mimic, while the mutation of predicted seed sequences of TGFBR3 3’-UTR partially restored the luminescence. Conclusions: The miR-193b may inhibit chondrogenesis of ATDC5 via targeting TGFBR3.View Large Image Figure ViewerDownload Hi-res image Download (PPT)View Large Image Figure ViewerDownload Hi-res image Download (PPT)