Abstract
Background/objectives: miR-21 has been linked to renal fibrosis and miR-155 too has shown different expression in IgAN whose pathogenesis is partially understood. In this study, we explored the utility of miR-21 and miR-155 in IgA nephropathy. Methods: We recruited 40 IgA nephropathy (IgAN) patients and 15 healthy controls (HC). 2 ml blood sample was collected from each study participant. Mean expressed value normalization and absolute and relative quantification methods were used for quantitative polymerase chain reaction (PCR) experiment for miRNA expression. Logistic regression analysis was performed for diagnostic and prognostic value of miR-21. Findings: Mean expressed value normalization reduced the variation in groups and between groups without changing the mean value. miR-155 showed similar expression in IgAN and healthy controls. There was three-fold increases in miR-21 in IgAN than the HC. miR-21 was able to differentiate the mesangial hypercellularity and tubular atrophy/interstitial fibrosis stages with sensitivity 84 and 70 percent, respectively. Applications: miR-155 can be used as a reference control gene for IgAN on quantitative PCR-based experiments. miR-21 can be used as a diagnostic and prognostic marker for IgAN. Keywords: miR-21, miR-155, IgAN, Tubular Atrophy, Interstitial Fibrosis.
Highlights
Introduction microRNAs are small RNA molecules of 18 to 22 nucleotides in length which play an important role in protein formation by interacting with mRNAs post transcriptionally [1]
[2] one miRNA can interact with multiple mRNA and vice versa and the presence, absence, increased, or decreased level of miRNA in any patient sample could be a sign of specific disease condition [3]
It starts with sample collection, miRNA isolation, complementary DNA conversion, and amplification stage
Summary
MicroRNAs (miRNAs) are small RNA molecules of 18 to 22 nucleotides in length which play an important role in protein formation by interacting with mRNAs post transcriptionally [1]. There are multiple stages involved in real-time PCR-based miRNA expression It starts with sample (fluid or/and tissue) collection, miRNA isolation, complementary DNA (cDNA) conversion, and amplification stage. Normalization techniques are used in real-time PCRbased miRNA expression experiments to reduce the technical variations for better and accurate measurement of biological variations of the disease [5] Mean expression value is frequently used for the normalization process of miRNA expression based studies [6]. This method is used to find the best suitable endogenous reference control for miRNA expression. There is different opinion to use exogenous miRNA as internal control in normalization process [7,8]
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