Development of Insulin Resistance through Induction of miRNA-135 in C2C12 Cells.
OBJECTIVE
Micro-RNAs (miRNAs) are a class of posttranscriptional 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.


METHODS
This experimental study was conducted on the C2C12 cell line. Using bioinformatics tools miRNA-135 and two respective target genes-insulin receptor (Insr) and vesicle associated membrane protein 2 (Vamp2)were selected as potential factors involved in insulin resistance process. Levels of glucose uptake miRNA expression and respective gene targets were determined after cell transfaction by miR-135.


RESULTS
It was determined that Insr gene expression was significantly down-regulated in miR-135 transfected C2C12 cell line (P≤0.05). Interestingly; these transfected cells have shown a significant difference in glucose uptake incomparision the positive control cells, while it was similar to the insulin resistant cell line (P≤0.05). In contrast, no significant alteration of Vamp2 gene expression was observed.


CONCLUSIONS
Our data indicated no change on the Vamp2 expression level after miRNA transfection, while expression level of Insr was reduced and miR-135 expression was contrarily increased leading to poor stimulation of glucose uptake through insulin, and development of insulin resistance phenotype in C2C12 cell line.
Department of Molecular Medicine, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran; Stem Cell Technology Research Center, Tehran, Iran.
Maryam
M
Honardoost
Department of Microbiology, School of Biology, College of Science, University of Tehran, Tehran, Iran.
Ehsan
E
Arefian
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Masoud
M
Soleimani
Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Sara
S
Soudi
Department of Molecular Medicine, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.
Mohammad Reza
MR
Sarookhani
C2C12; Insulin Receptor; Insulin Resistance; MiR-135