背景：索拉非尼是晚期肝细胞癌（HCC）的标准治疗方法，但治疗过程中的获得性耐药极大地限制了其临床疗效。脂质代谢紊乱在肝癌发生中起着重要作用。然而，脂质代谢重编程是否以及如何调节肝癌细胞的索拉非尼耐药性仍不清楚。方法：通过连续诱导建立索拉非尼耐药HCC细胞。 UHPLC-MS/MS、蛋白质组学和流式细胞术用于评估脂质代谢。 ChIP 和蛋白质印迹用于反映信号转导子和转录激活子 3 (STAT3) 与甘油-3-磷酸酰基转移酶 3 (GPAT3) 的相互作用。应用功能获得和丧失研究来探索 HCC 索拉非尼耐药的驱动机制。采用体外流式细胞术和CCK8以及体内肿瘤大小来评估HCC细胞对索拉非尼的敏感性。结果：我们的代谢组数据显示，索拉非尼耐药的 HCC 细胞中甘油三酯显着富集。使用蛋白质组学和基因组学技术的进一步分析表明，索拉非尼耐药组中 GPAT3 的表达显着增加，这依赖于 STAT3 的激活。 GPAT3的恢复使HCC细胞对索拉非尼重新敏感，而GPAT3的过度表达导致对索拉非尼不敏感。从机制上讲，GPAT3上调增加了甘油三酯的合成，进而刺激NF-κB/Bcl2信号通路，导致索拉非尼治疗后的细胞凋亡耐受。此外，我们的体外和体内研究表明，泛 GPAT 抑制剂可有效逆转 HCC 细胞中的索拉非尼耐药性。结论：我们的数据表明，HCC 细胞中 GPAT3 的升高会重新编程甘油三酯代谢，从而导致对索拉非尼的获得性耐药，这表明 GPAT3 是增强 HCC 对索拉非尼敏感性的潜在靶标。© 作者。

Background: Sorafenib is the standard treatment for advanced hepatocellular carcinoma (HCC), but acquired resistance during the treatment greatly limits its clinical efficiency. Lipid metabolic disorder plays an important role in hepatocarcinogenesis. However, whether and how lipid metabolic reprogramming regulates sorafenib resistance of HCC cells remains vague. Methods: Sorafenib resistant HCC cells were established by continuous induction. UHPLC-MS/MS, proteomics, and flow cytometry were used to assess the lipid metabolism. ChIP and western blot were used to reflect the interaction of signal transducer and activator of transcription 3 (STAT3) with glycerol-3-phosphate acyltransferase 3 (GPAT3). Gain- and loss-of function studies were applied to explore the mechanism driving sorafenib resistance of HCC. Flow cytometry and CCK8 in vitro, and tumor size in vivo were used to evaluate the sorafenib sensitivity of HCC cells. Results: Our metabolome data revealed a significant enrichment of triglycerides in sorafenib-resistant HCC cells. Further analysis using proteomics and genomics techniques demonstrated a significant increase in the expression of GPAT3 in the sorafenib-resistant groups, which was found to be dependent on the activation of STAT3. The restoration of GPAT3 resensitized HCC cells to sorafenib, while overexpression of GPAT3 led to insensitivity to sorafenib. Mechanistically, GPAT3 upregulation increased triglyceride synthesis, which in turn stimulated the NF-κB/Bcl2 signaling pathway, resulting in apoptosis tolerance upon sorafenib treatment. Furthermore, our in vitro and in vivo studies revealed that pan-GPAT inhibitors effectively reversed sorafenib resistance in HCC cells. Conclusions: Our data demonstrate that GPAT3 elevation in HCC cells reprograms triglyceride metabolism which contributes to acquired resistance to sorafenib, which suggests GPAT3 as a potential target for enhancing the sensitivity of HCC to sorafenib.© The author(s).