Serum miRNA-101 expression signature as non-invasive diagnostic biomarker for Hepatitis C virus-associated hepatocellular carcinoma in Egyptian patients

Abstract

Hepatocellular carcinoma (HCC) is a leading cause of cancer mortality globally due to HCC late diagnosis and limited treatment options. MiRNAs (miRNAs) emerged as potential biomarkers for various diseases, including HCC. However, the value of miRNA-101 as a serum biomarker for HCV-induced HCC has not been fully investigated. Our study aims to investigate the miRNA-101 differential expression in Egyptian HCV-induced HCC patients' serum versus HCV liver cirrhosis (LC) as prospective diagnostic biomarkers compared to alpha-fetoprotein (AFP). Blood samples were collected for clinical chemistry profile, liver function, and serum AFP investigations. The serum miR-101 expression levels were evaluated using real-time quantitative PCR (RT-qPCR) in 100 Egyptian subjects: 40 HCV-induced HCC, 40 HCV-induced cirrhosis, and 20 healthy controls. HCC patients showed significantly higher TB, DB, and AFP levels than those cirrhosis and control groups, whereas ALB and Total Protein exhibited significantly reduced levels. AFP sensitivity and specificity in differentiating HCC reported 60 and 67%, respectively, at the cut-off values of 7ng/dl. miR-101 shows fold change upregulation in HCC patients (P < 0.0001) compared to LC and control groups. ROC curve demonstrated miR-101 (AUC) of 0.9556, sensitivity 92.5%, and specificity 97.5%, highlighting the miR-101 diagnostic potential as a biomarker for HCC detection. Elevated miR-101 levels in HCC are significantly correlated with a higher number and larger size of focal lesions, advanced BCLC staging, and Child-Pugh score. These findings highlight the utility of miR-101 as a predictive and diagnostic non-invasive biomarker for HCV-related HCC from cirrhotic populations. More research is warranted to validate the clinical validity of miR-101 and explore underlying mechanisms in HCV-HCC progression.

Keywords: AFP; Hepatitis C virus (HCV); Hepatocellular carcinoma (HCC); Noninvasive Diagnostic Biomarkers; Therapeutic Targets; miRNA-101 expression.

Fig. 1

AFP Marker statistical analysis across study groups and control. Three separate replicates’ mean ± SE are displayed for each set of data. P. Value significant levels < 0.05 (low (*), < 0.01 Moderate (**), The unpaired t-test used for significance analysis between the groups.

Fig. 2

Triphasic CT images showed Large left lobe hepatic focal lesion (A) showed arterial enhancement (B) with rapid washout in delayed phase (C).

Fig. 3

miR-101 marker mean fold change values for differentiating HCV- HCC from HCV-liver cirrhotic group and control. Three separate replicates’ mean ± SE are displayed for each set of data. P. Value significant levels > 0.05 (ns), < 0.0001 high (****), The unpaired t-test used for significance analysis between the groups.

Fig. 4

Serum AFP- ROC curve analysis for distinguishing HCV- HCC group from the HCV-liver cirrhotic and control groups.

Fig. 5

ROC curve analysis of miR-101 as an HCV- HCC tumor marker.