Zhao Yanan, Li Kang, Sun Jianping, He Ning, Zhao Peng, Zang Chaoran, Yang Xiaozhen, Hu Caixia, Long Jiang, Zhang Honghai, Wang Qi, Zhao Yan, Zhang Yonghong
Research Center for Biomedical Resources, Beijing You'an Hospital, Capital Medical University, Beijing 100069, P.R. China.
Interventional Therapy Center for Oncology, Beijing You'an Hospital, Capital Medical University, Beijing 100069, P.R. China.
Oncol Lett. 2020 Jul;20(1):677-684. doi: 10.3892/ol.2020.11636. Epub 2020 May 18.
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.
乙型肝炎病毒（HBV）相关肝细胞癌（HCC）是中国最常见的肝癌类型。热消融是肝癌治疗的主要策略之一。然而，迄今为止，很少有研究调查热消融后免疫反应的特性。在本研究中，从北京佑安医院招募了5名HBV相关HCC患者。在三个时间点采集外周血单个核细胞（PBMC）：热消融前（PR）、消融后1 - 3天（P1）和消融后5 - 7天（P7）。采用Illumina 850K甲基化微阵列来确定每个样本的DNA甲基化谱。使用R语言中的Bioconductor软件包对不同的甲基化探针进行数据分析。在对不同的甲基化CG位点（CGs）进行注释后，对相关基因进行了Ingenuity通路分析。在PR、P1和P7时间点共鉴定出3000个显著不同的CGs（校正P<0.05；|log（倍数变化）|>0.5）。其中，744个（24.8%）位点在PR和P1时间点之间增加，但在P7时间点逐渐减少。其余2256个（75.2%）位点在PR和P1时间点之间减少，在P7时间点逐渐增加。在对启动子上不同CGs进行基因注释后，信号通路分析表明，“p70S6K信号通路”、“CXCR4信号通路”、“树突状细胞成熟”、“巨噬细胞中一氧化氮和活性氧的产生”通路在P7时间点被激活。本研究表明，对于HBV相关HCC患者，热消融后PBMC DNA甲基化很快发生变化，全身免疫反应被激活，特别是在P7时间点。
