Circulating microRNA-122 level in chronic hepatitis C with and without hepatocellular carcinoma in Egyptian patients

Abstract

Many studies demonstrate importance of miRNA 122 for effective viral translation and replication of HCV in vitro. This unique interaction may explain the increase of its level during the natural history of the virus in hepatocyte (Jopling et al. 2005). Recently, circulatory microRNAs were found to be a good biomarker for tumors and certain diseases due to their stability in the blood unlike long strand RNAs (Elfimova et al. 2012), among them miRNA 122 was found to be one of the few tissue specific miRNAs that are not affected by sample processing methods (Zhang et al. 2014). The aim of this study was to investigate for the difference of expression levels of miR-122 in cirrhotic HCV population with and without HCC in Egyptian patients. 40 patients were recruited from the Department of Internal Medicine and the Outpatient Clinic of Ain Shams University hospital in the period between June 2015 and June 2016 and 10 healthy volunteers representing the control. The patients were grouped as follow: 1- Group 1 (HCV group): composed of 20 patients with hepatitis C Virus defined by presence of HCV-RNA by Real-time PCR and no hepatic focal lesions (HFL) 2-Group 2 (HCC group): composed of 20 patients with known HCV who developed HCC which was diagnosed in accordance to recent guidelines. 138 Summary and conclusion 3-Group 3 (Control group): composed of 10 apparently healthy volunteers. -Exclusion of other causes of chronic liver disease including hepatitis B virus infection, self-reported alcohol consumption in the preceding six months, or autoimmune hepatitis, with ruling out the history of previous HCC treatment or antiviral therapy for HCV, with exclusion of presence of hepatic malignancies other than HCC. The patients were studied as follow: 1) Biochemical studies: Routine laboratory assessment (CBC, ALT, AST, Total and direct bilirubin, serum albumin, creatinine, BUN, PT PTT) and Alpha feto protein. 2) Radiological studies: All patients were submitted to ultrasound examination for assessment of cirrhosis, ascites and focal lesions. Patients with HFL underwent further characterization using either dynamic MSCT contrast enhanced scanning and/or dynamic contrast enhanced MRI scanning, where patients with typical imaging criteria for HCC were included into the corresponding group. 3) miRNA 122 expression BY Total RNA extraction and Reverse transcription and TaqMan real-time PCR assays of microRNA 139 Summary and conclusion Statistical Analysis: Data were analyzed on an IBM personal computer, using IBM Statistical Package for Special Science (SPSS) software, version 21. Data were described as mean ± standard deviation (SD) for quantitative (numerical) variables and as median & interquartile range (IQR) (categorical) variables. Non-parametric data were compared using Kruskal-Wallis test followed by Mann-Whitney post-hoc test for pair-wise analysis. Kolmogorov-Smirnov Z test was used for comparing the test to control groups (pair-wise analysis). Parametric data were compared using the Analysis Of Variance (one-way ANOVA) test followed by the LSD post-hoc test for pair-wise analysis Correlation of non-parametric data was done using the spearman’s correlation test. Significance level (P) value: P ≤ 0.05 is significant (S), P > 0.05 is insignificant (NS). The results were then tabulated and presented in box and whisker plots and pie charts. We found significant fold decrease of plasma miR-122 in both HCV & HCC groups as a whole than control but there were significant increase of plasma miR-122 in compensated HCV liver cirrhosis than control with subsequent significant decrease than control of its levels in decompensated HCV liver cirrhosis.