Identification of Circulating miRNAs as Non-Invasive Biomarkers of Triple Negative Breast Cancer in the Population of Pakistan
Identification of Circulating miRNAs as Non-Invasive Biomarkers of Triple Negative Breast Cancer in the Population of Pakistan
Jawaria Shaheen1, Samiah Shahid1, Sabahat Shahzadi2, M. Waheed Akhtar1,3 and Saima Sadaf1,*
ABSTRACT
Triple negative breast cancer is the heterogeneous and most aggressive known subtype of breast cancer due to absence of estrogen, progesterone and HER2 receptors. We aimed to identify a panel of circulating miRNAs which can act as diagnostic and prognostic biomarkers specifically for TNBC. The blood samples were collected from 37 TNBC diagnosed patients and 34 age-matched non-TNBC controls to investigate the expression of miRNA panel. Plasma was isolated by standard protocol of centrifugation and then isolation of total RNA including miRNA was done. About 100ng of isolated miRNA was converted to cDNA after addition of poly (A) tailing and then expression profiling was done by using real time-qPCR technique in which miR-specific DNA primers were used to increase sensitivity of reaction. Expression of miRNA panel was normalized with mir-16 and expression fold change was calculated by 2-ΔΔCt method. Further analysis was done by SPSS, medcalc and Graphpad prism software. Area under curve analysis was used to get diagnosis outcome of selected miRNA panel. Upregulation of miRNA panel (mir-376c, mir-155, mir-17a and mir-10b) has been seen in the plasma samples of TNBC patient´s samples relative to non-TNBC control samples. The scatter plots and other statistical analysis have given differences in expression of four miRNA panel with significant p-value (p< 0.001), respectively. Kruskal Wallis test and Mann Whitney U test have clearly showed significance and association of miRNA panel with clinicopathological characteristics of breast cancer which have revealed their role as diagnostic biomarkers. AUC curve analysis also remarks their high sensitivity and specificity. We have identified panel of four miRNAs which can act as circulating diagnostic and prognostic biomarkers in triple negative breast cancer in the population of Pakistan. Especially, mir-376c, mir-155 and mir-17a may act as early stage biomarkers in TNBC and mir-10b is found to be late stage biomarker which can also act as prognostic biomarker in TNBC.
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