通过诱导C2C12细胞中的miRNA-135产生胰岛素抵抗

Development of Insulin Resistance through Induction of miRNA-135 in C2C12 Cells.

作者信息

Honardoost Maryam, Arefian Ehsan, Soleimani Masoud, Soudi Sara, Sarookhani Mohammad Reza

机构信息

Department of Molecular Medicine, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran; Stem Cell Technology Research Center, Tehran, Iran.

Department of Microbiology, School of Biology, College of Science, University of Tehran, Tehran, Iran.

出版信息

Cell J. 2016 Fall;18(3):353-61. doi: 10.22074/cellj.2016.4563. Epub 2016 Aug 24.

Abstract

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.

MATERIALS AND 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.

CONCLUSION

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.

摘要

目的

微小RNA(miRNA)是一类转录后调节因子,在各种生物学过程中发挥关键作用。新出现的证据表明,miRNA与包括2型糖尿病(T2D)在内的几种疾病的发生发展存在直接联系。在本研究中,我们旨在探讨预测的miRNA及其靶基因对胰岛素抵抗的影响。

材料与方法

本实验研究在C2C12细胞系上进行。使用生物信息学工具,选择miRNA-135及其两个相应的靶基因——胰岛素受体(Insr)和囊泡相关膜蛋白2(Vamp2)作为参与胰岛素抵抗过程的潜在因素。在通过miR-135进行细胞转染后,测定葡萄糖摄取、miRNA表达水平及相应基因靶点。

结果

发现在转染miR-135的C2C12细胞系中,Insr基因表达显著下调(P≤0.05)。有趣的是,与阳性对照细胞相比,这些转染细胞在葡萄糖摄取方面显示出显著差异,而与胰岛素抵抗细胞系相似(P≤0.05)。相比之下,未观察到Vamp2基因表达有明显变化。

结论

我们的数据表明,miRNA转染后Vamp2表达水平无变化,而Insr表达水平降低,miR-135表达水平反而升高,导致胰岛素对葡萄糖摄取的刺激作用减弱,C2C12细胞系出现胰岛素抵抗表型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f5b1/5011323/b00a5bfa276b/Cell-J-18-353-g01.jpg

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