Circulating microRNAs as biomarkers for diagnosis of early hepatocellular carcinoma associated with hepatitis B virus

Abstract

Hepatocarcinogenesis is a multistep process that evolves from cirrhosis or dysplastic nodule (DN), and eventually leads to overt hepatocellular carcinoma (HCC). Differentiation between early HCC and DN is an important issue in the clinical setting. This study aims to investigate the potential of circulating microRNA (miRNA) levels in the diagnosis of early HCC. RNA was extracted from sera of 30 chronic hepatitis B patients with pathologically proven DN and 120 age-and sex-matched patients with early HCC. Paired samples were collected from ten patients with DN who developed overt HCC in the follow-up. A panel of ten cancer-associated miRNAs was analyzed by quantitative real-time reverse-transcription polymerase chain reaction. Serum levels of miR-16, miR-122, miR-221, let-7b and miR-15b were significantly lower in patients with DN than in the HCC group. When DN progressed to overt HCC, serum miR-122, miR-let-7b and miR-15b levels increased significantly (p 5 0.046, 0.043 and 0.044, respectively). As a single marker, a-fetoprotein (AFP) and miR-122 as well as let-7b had the similar performance for differentiate HCC from DN. As limited to subjects with normal AFP, let-7b resulted in a sensitivity of 84.8% and a specificity of 50% in separating HCC and DN with a cutoff value of 3.5 (p 5 0.001). In conclusion, miR-122 and let-7b, which are upregulated in the serum of early-HCC patients, can be useful markers for differentiating early HCC from DN in chronic hepatitis B patients.Hepatocellular carcinoma (HCC) is the sixth most common malignancy, and the third leading cause of cancer-related death in the world.1 Approximately 80-90% of patients with HCC have the underlying viral etiologies. 2-4 From a global perspective, the most important risk factor for HCC is chronic hepatitis B virus (HBV) infection, which is highly endemic in many Asian countries including Taiwan. 2-5 The overall 5-year survival rate is 5-9% from the time of clinical diagnosis of HCC, and the dismal prognosis is largely caused by late detection of the tumors. 6,7 Therefore, early detection of HCC at a curative stage offers the best chance of survival for these patients. Currently, diagnosis of HCC is based on imaging studies and sometimes verified by biopsy results.8 Advances in magnetic resonance imaging (MRI) and computed tomography (CT) have greatly improved imaging of focal hypervascular masses consistent with HCC, but these procedures are costly and not suitable for daily practice. Thus, sensitive and specific cancer biomarkers are essential for early detection and diagnosis of HCC as well as for developing preventive screening. Laboratory analyses including serum a-fetoprotein (AFP) and des-gamma carboxyprothrombin levels have long been used as HCC biomarkers. However, the accuracy of AFP is modest (sensitivity: 39-65%; specificity: 76-94%) and onethird of cases of early-stage HCC are missed using AFP analysis. 9 Also, serum AFP levels are probably false positive in patients with benign liver diseases, such as hepatitis and cirrhosis.10

Biomarker MicroRNAs for Diagnosis, Prognosis and Treatment of Hepatocellular Carcinoma: A Functional Survey and Comparison

“…Resisting cell death was significantly associated with lower expression of miR-101-3p, miR-16-5p, miR-195-5p, miR-203a-3p and miR-221-3p in blood samples38414243. Increased miR-221-3p, miR-224-5p, miR-483-5p and miR-122-5p expression were also detected in blood of HCC patients4044. These above diagnostic biomarkers as classifiers for HCC detection, yielding a receiver operating characteristic curve area of 0.635 to 0.884 (see Tables 1 and 2).…”

“…High expression of miR-106b-5p, miR-21-5p, miR-331-3p and low expression of miR-101-3p, miR-125b-5p, miR-139-5p had great potential to be noninvasive and accurate circulating biomarkers for HCC preliminary screening103851555661. Moreover, some opposite results about the expression levels of miR-122-5p were discussed4486. In tissues, high expression of eight miRNAs ( i.e.…”

Circulating microRNA profiles in plasma: identification of miR-224 as a novel diagnostic biomarker in hepatocellular carcinoma independent of hepatic function

“…Indeed, there was no significant correlation between the plasma miR-224 levels and various indicators of hepatic function, such as platelet, albumin, T-Bil, Alb, PT (%), AST, ALT and ICG 15R ( Figure 6 and Supplementary Figure S3B). In clinical settings, conventional tumor markers lack sufficient sensitivity and specificity to facilitate HCC detection, and previous reported biomarkers for HCC often reflect background liver disease and hepatic function [76][77][78][79][80]. Whereas, the effectiveness of plasma miR-224 was demonstrated to facilitate the sensitive HCC detection of smaller tumors of less than 18 mm (AUC: 0.802).…”

Non-coding RNAs: New therapeutic targets and opportunities for hepatocellular carcinoma

“…In the serum of patients with HCV-related HCC, low specific expressions of miR-30c-5p, miR-223-3p, miR302c-5p and miR-17-5p, as well as high specific expression of miR-221, indicated the recurrence of HCC [87,88] .…”