Evaluation of miR-331-3p and miR-23b-3p as serum biomarkers for hepatitis c virus-related hepatocellular carcinoma at early stage

Highlights

• Higher levels of serum miR-331-3p were demonstrated in early-stage HCC patients.

• Lower levels of serum miR-23b-3p were demonstrated in early-stage HCC patients.

• Serum levels of the two miRNAs were negatively correlated with liver fibrosis.

• Serum miR-331-3p and miR-23b-3p are potential biomarkes in HCC diagnosis.

Summary

Background and aims

This study aimed to investigate the diagnostic values of serum miR-331-3p and miR-23b-3p as tumor markers for the diagnosis of hepatocellular carcinoma (HCC) at early stage.

Methods

A total of 191 subjects were enrolled and consisted of 45 healthy controls (HC), 106 hepatitis c virus (HCV)-related chronic liver disease (CLD) patients, and 40 early-stage HCC patients. CLD patients were subdivided according to Metavir fibrosis-scoring. Serum miR-331-3p and miR-23b-3p were measured. The area under curves (AUC) was calculated for each microRNA and compared with that for alpha-fetoprotein (AFP) in the detection of HCC at early stage.

Results

Serum miR-331-3p was significantly higher in early-stage HCC than that in CLD and HC respectively, and it decreased significantly after surgery in early-stage HCC. Contrarily, serum miR-23b-3p was significantly lower in early-stage HCC and increased significantly after surgery. Further, receiver operating characteristic analysis demonstrated AUC was 0.806 (95%CI: 0.728–0.883; sensitivity: 85.85%, specificity: 65.00%) for serum miR-23b-3p in discriminating early-stage HCC from CLD patients, higher than that for AFP (AUC:0.660, 95%CI: 0.556–0.764; sensitivity: 70.00%, specificity: 56.60%). In discrimination early-stage HCC from severe fibrosis/cirrhosis (F3 + F4) patients, both miR-23b-3p (AUC: 0.796, 95%CI: 0.703–0.889; sensitivity: 85.11%, specificity: 65.00%) and miR-331-3p (AUC:0.832, 95%CI: 0.812–0.953; sensitivity: 75.00%, specificity: 85.11%) had better diagnostic performances than AFP (AUC:0.632, 95%CI: 0.512-0.753; sensitivity: 50.00%, specificity: 55.32%). Serum miR-331-3p levels also showed a significant correlation with BCLC stages of HCC.

Conclusion

Serum miR-331-3p and miR-23b-3p could be used as novel invasive biomarkers in the early detection of HCC in high-risk patients.

Introduction

Hepatocellular carcinoma (HCC) is the most common primary liver cancer and the third leading cause of cancer-related death in the world [1], [2]. Hepatitis C virus (HCV) infection in one of the main risk factors for developing HCC.

While great progress has been made in HCC treatment, the HCC prognosis remains poor and side effects of chemotherapy in patients are common [1]. The result of treatment depends on the HCC stage at the time of diagnosis. Early treatment of HCC can greatly improve life expectancy and reduce mortality. Although alpha-fetoprotein (AFP) serves as the most widely used marker for screening HCC patients, its sensitivity is quite low [3], [4], which is not sufficient for accurate diagnosis. Therefore new tumor markers to complement AFP to distinguish the early stage of HCC from risk persons are urgently needed.

MicroRNAs (miRNAs) are small, noncoding regulatory RNAs, involved in various biological processes including proliferation, differentiation and apoptosis [5]. It is well established that miRNAs may act as tumor suppressors, oncogenes, or even possess a dual nature playing both roles depending on the cellular needs in tumor occurrence and development [6], [7]. Most miRNAs can be released to blood and miRNAs are stable in plasma or serum [8]. As such, circulating miRNAs have been proposed to be promising biomarkers for noninvasive diagnosis and monitoring of tumors including HCC [8], [9], [10], [11], [12].

MiR-331-3p is one of the most commonly upregulated miRNAs that promote proliferation, migration and invasion of the tumor cells, including pancreatic tumor, breast tumor and HCC [13], [14], [15]. MiR-23b-3p is a down-regulated miRNA in the tumor, suppressing tumor cells proliferation and migration [16], [17], [18]. Recent studies showed miR-331-3p and miR-23b-3p were aberrantly expressed in HCC tissues [13], [17], and our previous screened study also found miR-331-3p and miR-23b-3p were differently expressed in the serum of HCC patients versus liver cirrhosis controls. However, the values of miR-331-3p and miR-23b-3p in serum for diagnosis of HCC have not been fully evaluated. The aim of this study was to investigate the values of serum miR-331-3p and miR-23b-3p as potential biomarkers in the screening of HCV-related HCC at early stage and their relationships with liver fibrosis.