Assessment of MicroRNAs as Biomarkers of Hepatitis C Virus-associated Hepatocellular carcinoma in Egyptian Patients

Abstract

Hepatitis C virus (HCV) is one of the major risk factors for the development of HCC. MiRNAs are short (19-24) nucleotides in length, non-coding RNAs that regulate protein synthesis at the transcriptional and translational levels. MiRNAs have been shown to regulate a variety of cellular processes, and their dysregulation plays a key role in cancer development.The current study was initiated to detect the expression levels of the liver cancer-associated miRNAs namely miR-17, miR-21, miR-25, miR-30c, miR-93, miR-106b and miR-222 in sera of HCC and HCV-infected Egyptian patients using qRT-PCR technique. In addition, the study aimed to examine the correlation between the levels of these miRNAs and different clinico-pathological parameters. To find the concordance between the expression levels of serum and liver tissue miRNAs, these miRNAs were detected in 25 HCC liver tissue samples using qRT-PCR. This study included 3 different groups: (1) HCC group included 25 patients with HCV-related HCC; (2) HCV group included 52 patients and (3) control group that included 50 healthy individuals. SerummiR-21, miR-25, miR-30c and miR-222 were significantly up-regulated whereas miR-17 and miR-106b were found to be down- regulated in HCC cases compared to controls. In HCV serum samples, miR-21, miR-25, miR-93 and miR-222 were found to be over-expressed whereas miR-30c was down-expressed. In liver tissue samples, two miRNAs (miR-30c and miR-106b) showed down-regulation and their fold changes were 0.12 and 0.28 respectively, while two miRNAs (miR-93 and miR-222) showed up-regulation and their mean fold changes recorded were 6.96 and 6.88 respectively. MiR-21 didn't show any significant alteration (fold change was 0.84). Matching was observed in miR-106b and miR-222 expression levels between serum and tissue samples in HCC patients.No significant correlation was detected between the relative expression level of miR-106b and clinico-pathological parameters. In contrast, a significant correlation was observed between the expression level of miR-222 and liver cirrhosis (P= 0.048). In addition, a highly significant correlation (P<0.001) was detected between the expression levels of the two miRNAs. ROC curve has showed that the serum level of miR-106b has moderate sensitivity (46.2%) and high specificity (84%) while miR-222 has moderate sensitivity (72%) and specificity (59.6%) in HCC diagnosis. The present study showed that miR-106b and miR-222 were deregulated in both HCC- and HCV- samples and that both of them have some diagnostic potential for primary HCC in chronically HCV-infected patients. However, further studies are needed to evaluate the diagnostic power of circulating miR-106b and miR-222 for hepatocellular carcinoma.