Mechanism of miR-133a-3p Negatively Regulating NG2 in Myofibroblasts Activation
ZHANG Hang, YUE Wenhui, YANG Le, LI Liying*
The aim of this study is to explore miRNA (microRNA) regulation of NG2 (neuron-glial antigen 2, gene name: Cspg4) expression and its molecular mechanism in the process of activating MFs (myofibroblasts). Mouse BMSCs (bone marrow mesenchymal stromal cells) were transfected with miRNA mimics and induced with TGFβ1 (transforming growth factor β1) to differentiate into MFs. The expression of Cspg4 and MFs activation markers were measured by qRT-PCR. The binding of Cspg4 and miRNA was detected by dual luciferase reporter gene assay. MFs activation up-regulated Cspg4 expression in a time-dependent and dose-dependent manner. The mRNA expression of Cspg4 was positively correlated with MFs activation markers. The mRNA expression of MFs activation markers were down-regulated after Cspg4 siRNA transfection. In the mouse model of liver injury, the miR-133a-3p expression was down-regulated and negatively correlated with Cspg4. miR-133a-3p binded to specific sites in the Cspg4 3ʹ UTR (3ʹ untranslated region) for post-transcriptional regulation. TGFβ1-induced up-regulation of Cspg4 and MFs activation markers were inhibited by miR-133a-3p. In conclusion, miR-133a-3p is involved in the activation of MFs by negatively regulating the expression of NG2.
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