Objectives: Apatinib has been reported to be a promising treatment for sorafenib-resistant hepatocellular carcinoma (HCC) patients. However, the underlying mechanism remains ambiguous. The study aimed to explore the efficacy of apatinib in sorafenib-resistant HCC and the underlying mechanism both in vitro and in vivo.

Methods: After observing epithelial-mesenchymal transformation (EMT) changes in HepG2 and HepG2/Sorafenib cells, we treated them with varying concentrations of apatinib to assess its impact on sorafenib-resistant HCC. Subsequently, specific inhibitors of c-Jun N-terminal kinase (JNK, SP600125) and extracellular signal-regulated kinase (ERK, PD98059) were introduced to investigate whether apatinib influenced sorafenib-resistant HCC via modulation of the epidermal growth factor receptor (EGFR)/JNK/ERK signaling pathway in vitro and in vivo. Biological behavior changes were assessed through cell counting kit-8 (CCK-8), colony formation, transwell, and immunofluorescence tests. Simultaneously, Western blot analysis was conducted to elucidate the expression of proteins associated with EMT and the EGFR/JNK/ERK signaling pathway.

Results: The HepG2/Sorafenib cells exhibited greater resistance to sorafenib compared to HepG2 cells, and sorafenib-resistant HCC was characterized by EMT changes. Apatinib demonstrated concentration-dependent inhibition of biological behaviors in HepG2/Sorafenib cells, with minimal impact on HepG2 cells. Additionally, apatinib had a pronounced effect on the expression of EMT-related proteins in sorafenib-resistant cells similar to that in sorafenib-sensitive cells. Furthermore, there was a dose-dependent reduction in the expression of proteins associated with the EGFR/JNK/ERK pathway in apatinib-treated groups. Notably, SP600125 and PD98059 contributed to the inhibition of EMT and EGFR/JNK/ERK pathway-related proteins by apatinib in sorafenib-resistant HCC.

Conclusion: Apatinib potentially hindered the progression of sorafenib-resistant HCC by suppressing both EMT and the EGFR/JNK/ERK pathway.

Keywords: Apatinib; EGFR/JNK/ERK; Epithelial mesenchymal transformation; Hepatocellular carcinoma (HCC); Sorafenib resistance.

© 2025 The Authors.

目标： 阿帕替尼已被报道可能是索拉非尼耐药的肝细胞癌（HCC）患者的有希望的治疗方法。然而，其潜在机制仍不清楚。本研究旨在探讨阿帕替尼在体外和体内对索拉非尼耐药性HCC的有效性和潜在机制。

方法： 观察肝细胞癌（HepG2）及其索拉非尼耐药株（HepG2/Sorafenib）的上皮-间质转化（EMT）变化后，我们使用不同浓度的阿帕替尼来评估其对索拉非尼耐药性HCC的影响。随后引入c-Jun N末端激酶（JNK, SP600125）和细胞外信号调节激酶（ERK, PD98059）的特定抑制剂，研究阿帕替尼是否通过调节表皮生长因子受体（EGFR)/JNK/ERK信号通路影响索拉非尼耐药性HCC。通过细胞计数试剂盒-8 (CCK-8)、集落形成、transwell和免疫荧光试验评估生物行为变化。同时进行Western blot分析以阐明与EMT及EGFR/JNK/ERK信号通路相关的蛋白质表达。

结果： HepG2/Sorafenib细胞表现出比HepG2细胞更强的索拉非尼耐药性，且索拉非尼耐药性肝癌具有EMT变化特征。阿帕替尼对HepG2/Sorafenib细胞显示出浓度依赖性的生物行为抑制作用，并对HepG2细胞影响较小。此外，阿帕替尼在索拉非尼耐药细胞中的相关蛋白表达有显著影响，类似于其在索拉非尼敏感细胞中的效果。同时，在阿帕替尼治疗组中与EGFR/JNK/ERK通路相关的蛋白质表达呈剂量依赖性降低。值得注意的是，SP600125和PD98059有助于抑制EMT及EGFR/JNK/ERK路径相关蛋白的形成。

结论： 阿帕替尼可能通过抑制上皮-间质转化（EMT）和表皮生长因子受体（EGFR)/JNK/ERK通路，阻止索拉非尼耐药性肝细胞癌的发展。

关键词： 阿帕替尼；EGFR/JNK/ERK；上皮间质转化；肝细胞癌（HCC）；索拉非尼耐药