Wang Lude, Zhuang Bo, Jiang Yiwen, Chen Zewei, Ge Chenyang, Yu Min, Yu Shian, Lin Haiping
Key Laboratory of Nutrition and Metabolism Research for Oncology, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, Zhejiang, China; Central Laboratory and Precision Medicine Center, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, Zhejiang, China.
Department of General Surgery, Jinhua Central Hospital, Teaching Hospital of Mathematical Medicine College, Zhejiang Normal University, Zhejiang, China.
Cell Signal. 2025 Nov;135:112076. doi: 10.1016/j.cellsig.2025.112076. Epub 2025 Aug 19.
Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide. Although sorafenib is a first-line systemic therapy for advanced HCC, resistance to treatment remains a major challenge. Hypoxia is a hallmark of the tumor microenvironment and is known to promote tumor growth and progression; however, its role in modulating sorafenib response in HCC remains poorly understood.
HCC cell lines (Hep3B and Huh7) were cultured under normoxic or hypoxic conditions and treated with sorafenib. Cell viability, apoptosis, and colony formation assays were conducted to evaluate the effect of hypoxia on sorafenib sensitivity. Luciferase reporter and chromatin immunoprecipitation (ChIP) assays were used to confirm HIF1A-mediated transcriptional regulation. RNA sequencing, m5C-RIP-Seq, and bioinformatics analysis were performed to identify NSUN2-mediated downstream targets. The therapeutic efficacy of GDF15 neutralization in combination with sorafenib was assessed both in vitro and in vivo.
Hypoxia significantly reduced HCC cell sensitivity to sorafenib, as evidenced by increased IC50 values and decreased apoptosis. Mechanistically, hypoxia-induced HIF1A upregulated the RNA m5C methyltransferase NSUN2, which stabilized GDF15 mRNA through m5C modification, leading to enhanced GDF15 secretion. Neutralization of GDF15 inhibited Akt/mTOR signaling and enhanced sensitivity to sorafenib in both in vitro and in vivo models.
This study uncovers a novel mechanism by which hypoxia-induced HIF1A promotes sorafenib resistance in HCC via the NSUN2-mediated stabilization and upregulation of GDF15. Targeting the HIF1A/NSUN2/GDF15 axis offers a promising therapeutic strategy to overcome sorafenib resistance in HCC patients.
肝细胞癌（HCC）是全球癌症相关死亡的主要原因。尽管索拉非尼是晚期HCC的一线全身治疗药物，但对治疗的耐药性仍然是一个重大挑战。缺氧是肿瘤微环境的一个标志，已知其可促进肿瘤生长和进展；然而，其在调节HCC中索拉非尼反应的作用仍知之甚少。
将HCC细胞系（Hep3B和Huh7）在常氧或缺氧条件下培养，并用索拉非尼处理。进行细胞活力、凋亡和集落形成试验以评估缺氧对索拉非尼敏感性的影响。使用荧光素酶报告基因和染色质免疫沉淀（ChIP）试验来确认HIF1A介导的转录调控。进行RNA测序、m5C-RIP-Seq和生物信息学分析以鉴定NSUN2介导的下游靶点。在体外和体内评估GDF15中和联合索拉非尼的治疗效果。
缺氧显著降低了HCC细胞对索拉非尼的敏感性，IC50值升高和凋亡减少证明了这一点。机制上，缺氧诱导的HIF1A上调了RNA m5C甲基转移酶NSUN2，其通过m5C修饰稳定了GDF15 mRNA，导致GDF15分泌增加。GDF15的中和在体外和体内模型中均抑制了Akt/mTOR信号传导并增强了对索拉非尼的敏感性。
本研究揭示了一种新机制，即缺氧诱导的HIF1A通过NSUN2介导的GDF15稳定和上调促进HCC中的索拉非尼耐药性。靶向HIF1A/NSUN2/GDF15轴为克服HCC患者的索拉非尼耐药性提供了一种有前景的治疗策略。
