Serum MicroRNA-21 as Marker for Necroinflammation in Hepatitis C Patients with and without Hepatocellular Carcinoma

TLDR

MicroRNA‑21 is overexpressed in tumor tissue and detectable at elevated levels in the serum of patients with various malignancies, making it a promising non‑invasive cancer marker. This study aimed to determine whether serum miR‑21 levels correlate with clinical and histological indicators of liver disease in chronic hepatitis C patients, including those with HCV‑associated hepatocellular carcinoma. Prospectively, 62 CHC patients, 29 CHC‑HCC patients, and 19 healthy controls were enrolled; serum miR‑21 and miR‑16 were quantified by qRT‑PCR and correlated with standard liver biochemistry, histological grading, and staging. Serum miR‑21 was higher in CHC patients than controls but unchanged in HCC, correlated inversely with HAI, ALT, AST, bilirubin, INR, and GGT, and independently predicted HAI; at a dCT cut‑off of 1.96 it distinguished minimal from moderate‑severe necroinflammation (AUC 0.758, 53.3 % sensitivity, 95.2 % specificity), confirming its role as a necroinflammatory marker but not a discriminator of HCC.

Abstract

Background MicroRNA-21 (miR-21) is up-regulated in tumor tissue of patients with malignant diseases, including hepatocellular carcinoma (HCC). Elevated concentrations of miR-21 have also been found in sera or plasma from patients with malignancies, rendering it an interesting candidate as serum/plasma marker for malignancies. Here we correlated serum miR-21 levels with clinical parameters in patients with different stages of chronic hepatitis C virus infection (CHC) and CHC-associated HCC. Methodology/Principal Findings 62 CHC patients, 29 patients with CHC and HCC and 19 healthy controls were prospectively enrolled. RNA was extracted from the sera and miR-21 as well as miR-16 levels were analyzed by quantitative real-time PCR; miR-21 levels (normalized by miR-16) were correlated with standard liver parameters, histological grading and staging of CHC. The data show that serum levels of miR-21 were elevated in patients with CHC compared to healthy controls (P<0.001); there was no difference between serum miR-21 in patients with CHC and CHC-associated HCC. Serum miR-21 levels correlated with histological activity index (HAI) in the liver (r = −0.494, P = 0.00002), alanine aminotransferase (ALT) (r = −0.309, P = 0.007), aspartate aminotransferase (r = −0.495, P = 0.000007), bilirubin (r = −0.362, P = 0.002), international normalized ratio (r = −0.338, P = 0.034) and γ-glutamyltransferase (r = −0.244, P = 0.034). Multivariate analysis revealed that ALT and miR-21 serum levels were independently associated with HAI. At a cut-off dCT of 1.96, miR-21 discriminated between minimal and mild-severe necroinflammation (AUC = 0.758) with a sensitivity of 53.3% and a specificity of 95.2%. Conclusions/Significance The serum miR-21 level is a marker for necroinflammatory activity, but does not differ between patients with HCV and HCV-induced HCC.