Genomic DNA methylation profiling indicates immune response following thermal ablation treatment for HBV-associated hepatocellular carcinoma.

Abstract

Hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC) is the most common type of liver cancer in China. Thermal ablation is one of the main strategies for HCC treatment. However, few studies have investigated the properties of the immune response following thermal ablation thus far. In the present study, five subjects with HBV-associated HCC were recruited from The Beijing Youan Hospital. Peripheral blood mononuclear cells (PBMCs) were collected at three time points: Prior to thermal ablation (PR), 1–3 days post-ablation (P1) and 5–7 days post-ablation (P7). An Illumina 850K methylation microarray was employed to determine the DNA methylation profile of each sample. Data were analyzed using different methylation probes with the Bioconductor package in R. Following annotation of different methylation CG sites (CGs), the associated genes were subjected to an Ingenuity Pathway Analysis. A total of 3,000 significantly different CGs (adjusted P<0.05; |log(fold-change)|>0.5) were identified within the PR, P1 and P7 time points. Of these, 744 (24.8%) sites increased between the PR and P1 time points but gradually decreased at the P7 time point. The remaining 2,256 (75.2%) sites decreased between the PR and P1 time points gradually increased at the P7 time point. Following gene annotation of different CGs on the promoter, signaling pathways analysis demonstrated that ‘p70S6K signaling’, ‘CXCR4 signaling’, ‘dendritic cell maturation’, ‘production of nitric oxide and reactive oxygen species in macrophages’ pathways were activated at the P7 time point. The present study suggested that PBMC DNA methylation had changed soon after thermal ablation for subjects with HBV-associated HCC, and systemic immune responses were activated, particularly at the P7 time point.