MiR‐199a‐3p inhibition facilitates cardiomyocyte differentiation of embryonic stem cell through promotion of MEF2C

Abstract

MicroRNAs (miRNAs) is a small molecule (19-25 nucleotide) noncoding RNA that inhibits the expression of target messenger RNA (mRNA) at the posttranscriptional level as an endogenous regulator. There is an increasing evidence that miR-199a-3p has a significant effect on the development of multiple tumors. However, the specific roles of miR-199a-3p in myocardial differentiation of embryonic stem cell still need to be investigated. Method of the hanging drop was used to build the model of cardiomyocyte differentiation of stem cell and beating rate of embryoid bodies (EBs) was calculated. The levels of intracellular MEF2C, a-MHC, GATA4, Nkx2.5, and cTnT mRNA were measured by real-time quantitative polymerase chain reaction, while the expressions of miR-199a-3p were detected simultaneously. Protein levels of MEF2C, a-MHC, GATA4, Nkx2.5, and cTnT were quantified by western blot analysis.Immunoreactivities of MEF2C and cTnT were analyzed by immunofluorescence.The interaction between miR-199a-3p and its predicted target (3′-untranslated region of MEF2C mRNA) was verified by luciferase assay. MiR-199a-3p levels increased during cardiogenesis. MiR-199a-3p inhibitor increased the beating rate of EBs and promoted expressions of cardiac-specific markers (GATA4, Nkx2.5, cTnT, and a-MHC). Notably, miR-199a-3p inhibition brought upregulation of MEF2C, which is the target of miR-199a-3p that we predicted and verified experimentally. In addition, MEF2C siRNA decreased miR-199a-3p inhibitor promoted EBs beating and attenuated miR-199a-3p inhibitor-induced cTnT and MEF2C expressions. The results above showed that MEF2C was involved in the process of promoting the differentiation of stem cells into cardiac myocytes by miR-199a-3p inhibitors. K E Y W O R D S cardiomyocyte, embryonic stem cell, miR-199a-3p, myocyte enhance factor 2C, myogenic How to cite this article: Chen H-P, Wen J, Tan S-R, Kang L-M, Zhu G-C. MiR-199a-3p inhibition facilitates cardiomyocyte differentiation of embryonic stem cell through promotion of MEF2C.

MicroRNA therapy confers anti-senescent effects on doxorubicin-related cardiotoxicity by intracellular and paracrine signaling

“…MiR-199a-3p, same as other microRNAs, engages a broad series of mRNA targets, including numerous survival factors and cell cycle regulatory proteins to take effects [ 36 , 37 ]. Thus, the potential promotion of the regenerative response by the miR-199a-3p involves the actions of protein targets that participate in cellular senescence [ 14 ].…”

PECAM1 Combines With CXCR4 to Trigger Inflammatory Cell Infiltration and Pulpitis Progression Through Activating the NF-κB Signaling Pathway

“…Importantly, the above bioinformatics analyses suggested MEF2C owned an over 0.6 correlation coefficient with both PECAM1 and CXCR4. As a subtype of the MEF2 protein family which are expressed in a variety of tissues, MEF2C was reported to play a crucial role in myocardiogenesis (Chen et al, 2019). Intriguingly, this transcription factor has been reported to be specifically activated by the MAP kinase p38 during inflammation (Han et al, 1997).…”

Effects of miRNAs, lncRNAs and circRNAs on osteoporosis as regulatory factors of bone homeostasis (Review)

“…a previous study highlighted the fact that mir-199a-3p plays a crucial role during adipocyte differentiation in ovariectomized mice (21). in addition, chen et al (22) revealed that mir-199a-3p participates in the inhibition of cardiomyocyte differentiation in stem cells via the MeF2c pathway. recently, Wu et al (23) found that mir-199a-3p expression was increased in ovariectomized mice and that the upregulation of mir-199a-3p inhibited the osteogenic differentiation of BMScs in vitro.…”