Lin Yang-Hsiang, Lai Ming-Wei, Chu Yu-De, Lin Kwang-Huei, Hsu Chao-Wei, Chien Rong-Nan, Chuang Po-Heng, Lin Chih-Lang, Yeh Chau-Ting
Liver Research Center, Chang Gung Memorial Hospital, Linkou. No. 15, Wenhua 1st Rd., Guishan Dist., Taoyuan City, 333, Taiwan.
Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
Cancer Cell Int. 2025 Jun 21;25(1):223. doi: 10.1186/s12935-025-03873-0.
A number of effective antiviral agents for chronic hepatitis B are available worldwide, and these agents have exhibited satisfactory virologic suppression, but suboptimal responses still occur in some patients. We aimed to explore the contribution of long non-coding RNAs (lncRNAs) in mediating suboptimal responses in chronic hepatitis B patients, which remains to be fully elucidated.
Cox regression models were used to analyze associations between suboptimal response and clinical factors. Hepatitis B virus X (HBx) gene was sequenced from entecavir-treated patients who developed hepatocellular carcinoma (HCC). Functional assays were conducted using Transwell assays, MTT assays, and xenograft model.
Suboptimal response was found to be a significant independent predictor of HCC development. Five of the six patients who developed HCC in the suboptimal response period were found to have HBx mutations (HBx-L100 insertion, HBx-G32R/K130M, HBx-Q87G/k130M/C143R, HBx-L123S, and HBx-H94Y/K130M). Overexpression of the HBx-H94Y/K130M mutation in HepG2.2.15 cells showed significantly increased cccDNA accumulation and enhanced cell migration compared to controls and other HBx mutants. RNA-seq analysis identified RPL13AP25 as a direct target of HBx-H94Y/K130M. RPL13AP25 was highly expressed in HCC tissues, and its elevated expression was associated with poor overall survival and enhanced cell motility and cccDNA accumulation both in vitro and in vivo. Mechanistically, both HBx-H94Y/K130M and RPL13AP25 enhanced the hyperphosphorylation of eIF4EBP1, leading to its dissociation from eIF4E, which subsequently enhances protein synthesis and ultimately contributes to HCC.
The HBx-H94Y/K130M mutant, selected during the period of suboptimal virological response, appears to promote cccDNA accumulation, likely through the upregulation of RPL13AP25, which contributed to HCC progression.
全球有多种用于慢性乙型肝炎的有效抗病毒药物，这些药物已显示出令人满意的病毒学抑制效果，但仍有一些患者出现疗效欠佳的情况。我们旨在探讨长链非编码RNA（lncRNA）在介导慢性乙型肝炎患者疗效欠佳反应中的作用，这一点仍有待充分阐明。
采用Cox回归模型分析疗效欠佳反应与临床因素之间的关联。对接受恩替卡韦治疗后发生肝细胞癌（HCC）的患者进行乙型肝炎病毒X（HBx）基因测序。使用Transwell实验、MTT实验和异种移植模型进行功能分析。
疗效欠佳反应被发现是HCC发生的一个显著独立预测因素。在疗效欠佳反应期发生HCC的6名患者中，有5名被发现存在HBx突变（HBx-L100插入、HBx-G32R/K130M、HBx-Q87G/k130M/C143R、HBx-L123S和HBx-H94Y/K130M）。与对照和其他HBx突变体相比，HepG2.2.15细胞中HBx-H94Y/K130M突变的过表达显示cccDNA积累显著增加且细胞迁移增强。RNA测序分析确定RPL13AP25是HBx-H94Y/K130M的直接靶点。RPL13AP25在HCC组织中高表达，其表达升高与总体生存率差以及体外和体内细胞运动性增强和cccDNA积累有关。从机制上讲，HBx-H94Y/K130M和RPL13AP25均增强了eIF4EBP1的过度磷酸化，导致其与eIF4E解离，随后增强蛋白质合成并最终促进HCC的发生。
在病毒学反应欠佳期选择的HBx-H94Y/K130M突变体似乎通过上调RPL13AP25促进cccDNA积累，这有助于HCC进展。
