Application of stem-loop RT-PCR procedure for evaluation of microRNA-21 in fresh tissues of invasive ductal carcinoma.

Background:- MiR-21 was one of the first oncogenic miRs and as an anti-apoptotic factor, to be characterized, being up-regulated in numerous tumors including BC . A novel miR quantification method has been established using stem–loop RT followed by TaqMan PCR analysis in tissues or culture cells. Aim of study:-Estimation of miR-21 expression level in fresh tissues of BC / NATs by using stem-loop follow by TagMan real time PCR (RT-PCR) technique and correlate miR-21 gene expression with clinico-pathological parameter of breast cancer . Material and methods:- Stem-loop RT-PCR was performed to identify the expression level of miR-21 in 50 IDC samples and their NATs. The expression levels of miR-21 relative to mRNA of GAPDH were determined using the livak method. Results:- Mean cancer tissue fold change of miR-21 was significantly higher than that of NATs. Majority of cases showed up regulation of miR-21, 96%. Up-regulation of miR-21 expression has been statistical significant associated with unfavorable pathological features of the disease, including positive lymph node involvement and higher tumor stage(III,VI) Conclusion:- Up-regulation of miR-21 expression has been associated with positive lymph node involvement and higher tumor stage(III,VI).