MicroRNA-96 impairs insulin signaling pathway by inhibiting IRS2 associated signaling molecules
Zeynab Hosseinpoor1 *, Zahra-Soheila Soheili2 , Maliheh Davari2 , Shahram Samie3 , Hamid Ahmadieh4
- National Institute of Genetic Engineering and Biotechnology
- National Institute of Genetic Engineering and Biotechnology
- Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
- Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Abstract: MicroRNAs have been recognized to play critical role in diabetic retinopathy (DR) development. We identified miR-96 as one of the most highly expressed miRNAs in diabetic retina. It subsequently regulates the expression level of its target genes, those involved in insulin signaling pathway in diabetic retina. We investigated the effects of miR-96 overexpression on its target genes in hRPE cell line.
Methods: miR-96 complete sequence was amplified from human genome through PCR. The amplified fragment was cloned into pAAV-MCS-eGFP-intron vector. hRPE cells were transfected with resultant construct. cDNA was synthesized and RT-qPCR was performed. The effects of miR-96 overexpression on growth and survival of transfected RPE cells were also investigated by MTT assay.
Results: The expression level of miR-96 was upregulated (more than 82-fold) in transfected hRPE cells. Moreover, the mRNA levels of IRS2, FOXO1 and ERK2 were significantly decreased in transfected hRPE cells. MTT assay revealed that miR-96 overexpression has no negative effect on growth and survival of hRPE cells.
Conclusion: Our results demonstrated that miR-96 overexpression might affect insulin pathway through different signaling molecules such as IRS2, FOXO1 and ERK2. In fact, miR-96 regulated the expression of target genes and controls biological processes. Taken together, modulation of miR-96 expression level may consider as a useful strategy for DR control.
