Association between overexpression of CDK9 and the expression of DNMTs gene in mouse myoblast C2C12 cells during differentiation.

Association between overexpression of CDK9 and the expression of DNMTs gene in mouse myoblast C2C12 cells during differentiation.
Leila Abkhooie,^1,* Mostafa Moradi Sarabi,² ‪Mohammad Saeid Hejazi‬,³
1. 1-Department of Molecular Medicine, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran & 2-Khorramabad, Department of Clinical Laboratory Sciences, School of Allied Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.
2. Department of Clinical Laboratory Sciences, School of Allied Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.
3. Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran
Introduction: Cdk9 is a member of cyclin-dependent kinases (cdks) that has been associated with cardiac differentiation. Increasing of Cdk9 expression level could moderate the epigenome. Also, DNA methyltransferases (DNMTs), responsible for the transfer of a methyl group from the universal methyl donor, S-adenosyl-L-methionine (SAM), to the 5-position of cytosine residues in DNA, are essential for mammalian development. There are four members of the DNMT family, including DNMT1, DNMT3A, DNMT3B, and DNMT3L. We proposed whether Cdk9 overexpression might affect the global genomic DNA methylation status by influencing DNMTs gene expression in mouse myoblast C2C12 cells during differentiation.
Methods: For overexpression of CDK9, the PCEP4/CDK9 plasmid was transfected to C2C12 cells. Transfected and control C2C12 cells were differentiated and the expression of DNA methyltransferase enzymes (DNMT1, 3A, and 3B) level was evaluated in transfected and control C2C12 cells by quantitative RT‐PCR upon Cdk9 overexpression.
Results: Our results showed that Cdk9 overexpression outcomes in DNA methylation changes in mouse myoblast C2C12 cells. We indicated that, upon Cdk9 overexpression, the average expression levels of DNMTs in proportion to global DNA methylation were meaningfully increased in Cdk9 transfected cells. Our study showed that Cdk9 caused to promoted epigenetic changes and moderate global DNA methylation profiling of the myoblast cells during differentiation.
Conclusion: In this study, we showed that although the expression of CDK9 enhances the cardiac differentiation in early stage of differentiation, however the overexpression of CDK9 cause to increase expression of DNA methyltransferase enzymes level during the cardiac differentiation. This change inhibits the differentiation by DNA methylation of myomiRs and genes promoters.
Keywords: DNA methyl-transferases, CDK9, C2C12 differentiation.