Methylation-sensitive High-Resolution Melting Analysis of the USP44 Promoter Can Detect Early-Stage Hepatocellular Carcinoma in Blood Samples.

Hepatocellular carcinoma (HCC) is dangerous cancer that often evades early detection because it is asymptomatic and an effec-tive detection method is lacking. For people with chronic liver inflammation who are at high risk of developing HCC, a sen-sitive detection method for HCC is needed. In a meta-analysis of The Cancer Genome Atlas pan-cancer methylation database, we identified a CpG island in the USP44 promoter that is me-thylated specifically in HCC. We developed methylation-sensi-tive high-resolution melting (MS-HRM) analysis to measure the methylation levels of the USP promoter in cell-free DNA isolated from patients. Our MS-HRM assay correctly identified 40% of patients with early-stage HCC, whereas the alpha-fetoprotein test, which is currently used to detect HCC, correctly identified only 25% of early-stage HCC patients. These results demonstrate that USP44 MS-HRM analysis is suitable for HCC surveillance.