Type 2 diabetes is one of the most prevalent diseases, which increases resistance to insulin in target tissues. The measurement of miRNAs quantity is a molecular approach for diagnosis of diabetes. miRNAs are small non-coding RNA strings of 21–23 long nucleotides that act as inhibitors in proteins translation. Several methods including Northern blot, qRT-PCR and Microarray have been used for diagnosis of miRNA molecules. Real time PCR is an expensive and accurate quantitative method that is widely used in miRNA studies. The miR-21 is an important miRNA in diabetes. In this study, for the first time, a semi-quantitative protocol was developed to quantify different amounts of a synthetic miR-21. In addition to semi-quantitative method, the miR-21 quantity was determined by quantitative method in several patients with type 2 diabetes and healthy people. The results indicated that there was a direct relationship between the amount of synthetic miR-21 and the intensity of the PCR bands. We also showed that the expression of miR-21 in people with type 2 diabetes increased compared to healthy people. The results were observed by both quantitative and semi-quantitative methods. The real-time RT-PCR was more sensitive than semi-quantitative PCR in identification of miRNAs. However, semi-quantitative PCR method benefited from higher simplicity and lower costs for defining general patterns of miRNA expression.
Type 2 diabetes miR-21 Semi-quantitative PCR Real-time RT-PCR
Phosphatase and tensin homolog
Tumor necrosis factor alpha
Interleukin 1 beta
This is a preview of subscription content, log in to check access.
This work was financially supported by graduate study of University of Kashan under Grant No. 572212/04.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This study was granted by the Aklaghezisti Ethical Committee at University of Kashan. All procedures were in accordance with the approved ethical standards of the Ethical Committee.
Written informed consent obtained from all healthy and diabetic people.
Guay C, Roggli E, Nesca V et al (2011) Diabetes mellitus, a microRNA-related disease? Transl Res 157(4):253–264CrossRefGoogle Scholar
de Planell-Saguer M, Rodicio MC (2013) Detection methods for microRNAs in clinic practice. Clin Biochem 46(10–11):869–878CrossRefGoogle Scholar
Zampetaki A et al (2010) Plasma microRNA profiling reveals loss of endothelial miR-126 and other microRNAs in type 2 diabetes. Circ Res 107:810–817CrossRefGoogle Scholar
Sekar D, Islam VIH, Thirugnanasambantham K et al (2014) Relevance of miR-21 in HIV and non-HIV-related lymphomas. Tumour Biol 35(9):8387–8393CrossRefGoogle Scholar
Dey N, Das F, Mariappan MM et al (2011) microRNA-21 orchestrates high glucose-induced signals to TORC1 for renal cell pathology in diabetes. J Biol Chem 286(29):68–75CrossRefGoogle Scholar
Roy S, Khanna S, Hussain SRA (2009) MicroRNA expression in response to murine myocardial infarction: miR-21 regulates fibroblast metalloprotease-2 via phosphatase and tensin homologue. Cardiovasc Res 82(1):21–29CrossRefGoogle Scholar
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucl Acids Res 29(9):e45CrossRefGoogle Scholar
Olivieri F, Rippo MR, Prattichizzo F et al (2013) Toll like receptor signaling in “inflammaging”: microRNA as new players. Immun Ageing 10(1):57–68CrossRefGoogle Scholar
Li S, Chen X, Zhang H et al (2009) Differential expression of microRNAs in mouse liver under aberrant energy metabolic status. J Lipid Res 50(9):1756–1765CrossRefGoogle Scholar
Meng S, Cao JT, Zhang B, Wang CQ et al (2012) Down regulation of microRNA-126 in endothelial progenitor cells from diabetes patients, impairs their functional properties, via target gene Spred-1. J Mol Cell Cardiol 53(1):64–72CrossRefGoogle Scholar