الفهرس 
Title : Investigating some micro-RNAs expression profiles in HCV Egyptian patients
AbstractOnly 14 pages are availabe for public view
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Abstract
Unresolved chronic hepatitis C virus (HCV) infection progresses into more deteriorating conditions such as fibrosis, cirrhosis and hepatocellular carcinoma (HCC). The latter is the fifth most common cancer worldwide and the third most common cause for cancer mortality. Currently, there is no reliable non-invasive biomarker for diagnosis, prognosis and staging of HCV mediated hepatic disorders. Extracellular miRNAs have emerged recently as promising non-invasive biomarkers in several diseases. The main objective for the current study is to profile expression signature for miR-147b, 512-5p, 552-3p, 650 & 676-3p in the plasma of 167 Egyptian patients affected with various stages of HCV associated disorders (40 healthy subjects, 48 HCV CLD (F0-F3) cases, 39 HCV cirrhotic cases and 40 HCV-HCC cases) using RT-PCR. miR-676-3p was then transfected into two different liver cancer cell lines (HepG2 and Huh-7) using lipofectamine 3000 reagent. The levels of miR-676-3p and mRNAs of the investigated genes were detected using RT-PCR. The investigated miRNAs were differentially expressed among all studied groups. All the investigated miRNAs except miR-147b could discriminate between HCC subjects and controls. Both miR-676-3p and miR-650 were powerful in discriminating cirrhotic and late fibrotic (F3-F4) cases from HCC. MiR-650 could distinguish mild (F0-F1) and advanced (F2-F3) CLD subjects from HCC cases. Both miR-650 and miR-147b could distinguish early fibrotic cases (F1-F2) from controls meanwhile miR-676 and miR-147b could effectively distinguish between mild CLD and (F1-F3) cases from healthy individuals. All studied miRNAs, except miR-512, can differentiate between (F0-F3) cases and controls. Multivariate logistic regression revealed three potential miRNA panels namely (miR-650 and miR-552-3p; miR-650 and miR-512-5p; miR-676-3p and miR-512-5p) for effective differentiation of HCC, cirrhotic and chronic liver cases respectively with high accuracy. Both miR-676-3p and miR-512-5p were significantly correlated in (F1-F3) fibrotic stage meanwhile miR-676 and miR-512-5p were found to differentiate between cirrhosis and (F0-F3) cases. Both miR-650 and miR-512-5p were positively correlated in the cirrhotic group and in (F0-F4) group .The results of in-vitro experiments demonstrated that transfection of miR-676-3p regulated the expression levels of several oncogenes and tumor suppressor genes in liver cancer cells. Notably, miR-676-3p transfection was found to downregulate Akt, β-catenin, TNF-α, Hsp90α and NFE2. Moreover, it significantly upregulated p53, GSK-3β and CASP7. On the other hand, the expression levels of Nox4, TGF-β and IL-6 were found unaltered in miR-676-3p transfected liver cancer cell lines. In conclusion, the expression signatures for the ` miRNAs could assist in staging and diagnosis with possible prognosis of various HCV associated hepatic disorders among Egyptian patients. Further large scale studies on other populations of various HCV genotypes are needed to investigate the possibility of using those miRNAs as universal biomarkers in HCV related liver diseases across the globe and that miR-676-3p may function as a novel tumor suppressor in HCC via regulating Akt and β-catenin signaling pathways, cytokine signaling, cancer-related inflammation and apoptosis. Accordingly, miR-676-3p could be a promising target for HCC therapy.