Mir-214 induces insulin resistance in c2c12 cell line though targeting rock1 and akt2 gene expressions

Maryam Honardoost,1 Maryam honardoost,2 Ehsan arefian,3 Masoud soleimani,4,*

1. Endocrine Research Center, Institute of Endocrinology and Metabolism, ran University of Medical Sciences, Tehran, Iran
2. Department of Microbiology, School of Biology, College of Science, University of Tehran, Tehran, Iran.
3. Department of Hematology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

Abstract

Introduction

Micro-rnas (mirnas) are a class of post-transcriptional regulators that play crucial roles in various biological processes. emerging evidence suggests a direct link between mirnas and development of several diseases including type 2 diabetes (t2d). in this study, we aimed to investigate the effect of predicted mirna and target genes on insulin resistance in order to the identification new potential biomarkers.

Methods

This experimental study was conducted on the c2c12 cell line. using bioinformatics tools mirna-214 and two respective target genes rho-associated coiled-coil containing protein kinase 1 (rock1) and akt serine/threonine kinase 2 (akt2)- were selected as potential factors involved in insulin resistance process. levels of glucose uptake mirna expression and respective gene targets were determined after cell transfection by mir-214. to check the specificity of the mirnas effect on targets luciferase assay were done.

Results

It was determined that rock1 and akt2 gene expression was significantly down-regulated in mir-214 transfected c2c12 cell line (p≤0.05). interestingly; these transfected cells have shown a significant difference in glucose uptake in comparison to the positive control cells, while it was similar to the insulin resistant cell line (p≤0.05). based on our results ,co-transfection of mir-214 with psicheck2-rock-1/ akt2 significantly decreased luciferase expression level. hence rock-1 and akt2 are direct the targets of mir 214.

Conclusion

Our data indicated expression level of rock1 and akt2 were reduced after mirna induction and mir-214 expression was contrarily increased leading to poor stimulation of glucose uptake through insulin, and development of insulin resistance phenotype in c2c12 cell line. to sum up, mir-214 may have a potential prognostic/diagnostic function in order to identify insulin resistance in type 2 diabetes.

Keywords

Mirna, insulin resistance