Liu Wen-Chun, Wu I-Chin, Lee Yen-Chien, Lin Chih-Peng, Cheng Ji-Hong, Lin Yih-Jyh, Yen Chia-Jui, Cheng Pin-Nan, Li Pei-Fu, Cheng Yi-Ting, Cheng Pei-Wen, Sun Koun-Tem, Yan Shu-Ling, Lin Jia-Jhen, Yang Jui-Chu, Chang Kung-Chao, Ho Cheng-Hsun, Tseng Vincent S, Chang Bill Chia-Han, Wu Jaw-Ching, Chang Ting-Tsung
Department of Internal Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan, ROC.
Infectious Disease and Signalling Research Centre, National Cheng Kung University, Tainan, Taiwan, ROC.
J Pathol. 2017 Oct;243(2):176-192. doi: 10.1002/path.4938. Epub 2017 Aug 17.
This study investigated hepatitis B virus (HBV) single-nucleotide variants (SNVs) and deletion mutations linked with hepatocellular carcinoma (HCC). Ninety-three HCC patients and 108 non-HCC patients were enrolled for HBV genome-wide next-generation sequencing (NGS) analysis. A systematic literature review and a meta-analysis were performed to validate NGS-defined HCC-associated SNVs and deletions. The experimental results identified 60 NGS-defined HCC-associated SNVs, including 41 novel SNVs, and their pathogenic frequencies. Each SNV was specific for either genotype B (n = 24) or genotype C (n = 34), except for nt53C, which was present in both genotypes. The pathogenic frequencies of these HCC-associated SNVs showed a distinct U-shaped distribution pattern. According to the meta-analysis and literature review, 167 HBV variants from 109 publications were categorized into four levels (A-D) of supporting evidence that they are associated with HCC. The proportion of NGS-defined HCC-associated SNVs among these HBV variants declined significantly from 75% of 12 HCC-associated variants by meta-analysis (Level A) to 0% of 10 HCC-unassociated variants by meta-analysis (Level D) (P < 0.0001). PreS deletions were significantly associated with HCC, in terms of deletion index, for both genotypes B (P = 0.030) and C (P = 0.049). For genotype C, preS deletions involving a specific fragment (nt2977-3013) were significantly associated with HCC (HCC versus non-HCC, 6/34 versus 0/32, P = 0.025). Meta-analysis of preS deletions showed significant association with HCC (summary odds ratio 3.0; 95% confidence interval 2.3-3.9). Transfection of Huh7 cells showed that all of the five novel NGS-defined HCC-associated SNVs in the small surface region influenced hepatocarcinogenesis pathways, including endoplasmic reticulum-stress and DNA repair systems, as shown by microarray, real-time polymerase chain reaction and western blot analysis. Their carcinogenic mechanisms are worthy of further research. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
本研究调查了与肝细胞癌（HCC）相关的乙型肝炎病毒（HBV）单核苷酸变异（SNV）和缺失突变。纳入了93例HCC患者和108例非HCC患者进行HBV全基因组二代测序（NGS）分析。进行了系统的文献综述和荟萃分析，以验证NGS定义的与HCC相关的SNV和缺失。实验结果确定了60个NGS定义的与HCC相关的SNV，包括41个新的SNV及其致病频率。除了在两种基因型中均存在的nt53C外，每个SNV对基因型B（n = 24）或基因型C（n = 34）具有特异性。这些与HCC相关的SNV的致病频率呈现出明显的U形分布模式。根据荟萃分析和文献综述，来自109篇出版物的167个HBV变异被分为与HCC相关的四个支持证据水平（A - D）。在这些HBV变异中，NGS定义的与HCC相关的SNV的比例从荟萃分析（A水平）中12个与HCC相关变异的75%显著下降到荟萃分析（D水平）中10个与HCC不相关变异的0%（P < 0.0001）。就缺失指数而言，前S区缺失与HCC显著相关，基因型B（P = 0.030）和基因型C（P = 0.049）均如此。对于基因型C，涉及特定片段（nt2977 - 3013）的前S区缺失与HCC显著相关（HCC与非HCC，6/34对0/32，P = 0.025）。前S区缺失的荟萃分析显示与HCC显著相关（汇总比值比3.0；95%置信区间2.3 - 3.9）。对Huh7细胞的转染显示，小表面区域中5个新的NGS定义的与HCC相关的SNV均影响肝癌发生途径，包括内质网应激和DNA修复系统，如微阵列、实时聚合酶链反应和蛋白质印迹分析所示。它们的致癌机制值得进一步研究。版权所有© 2017英国和爱尔兰病理学会。由John Wiley & Sons, Ltd.出版。
