Chen Xuechun, Ni Dongxuan, Cheng Jinghui, Liang Bin, Zhang Ruihan, Xiao Weilie, Liu Rong
Translational Cancer Research Center, Peking University First Hospital, Beijing, China.
Key Laboratory of Medicinal Chemistry for Natural Resource of Ministry of Education, Yunnan Characteristic Plant Extraction Laboratory Co., Ltd., Yunnan Research and Development Center for Natural Products, School of Life Sciences and School of Pharmacy, Yunnan University, Kunming, China.
Front Oncol. 2025 Sep 1;15:1641979. doi: 10.3389/fonc.2025.1641979. eCollection 2025.
Liver cancer remains a major global health burden, with hepatocellular carcinoma (HCC) accounting for approximately 80% of liver cancer cases. Cancer stem cells (CSCs) play a critical role in HCC initiation, progression, metastasis, and resistance to therapy, making them critical targets for novel therapeutic interventions. However, effective agents specifically targeting CSCs in HCC remain limited. The objective of this study was to identify and characterize novel small molecules that inhibit CSCs properties and overcome drug resistance in HCC.
Functional assays assessed the effects of C504244 on tumor sphere formation, cancer cell proliferation, and migration. RNA sequencing was conducted on C504244-treated HCC cells to investigate changes in gene expression profiles. Downstream targets of the Wnt signaling pathway were analyzed to determine pathway inhibition. Co-immunoprecipitation (Co-IP) was performed to assess whether C504244 disrupts the interaction between β-catenin and Transcription Factor 4 (TCF4) in HCC cells. Lenvatinib-resistant HCC cell lines were used to evaluate the combinatorial efficacy of C504244 and Lenvatinib and .
C504244 significantly suppressed tumor sphere formation, proliferation, and migration of HCC cells. Transcriptome analysis revealed that C504244 treatment led to significant inhibition of the Wnt signaling pathway, with corresponding downregulation of downstream target gene expression. Mechanistically, C504244 disrupted the β-catenin/TCF4 complex formation, which may contribute to reduced transcriptional activity. Since β-catenin signaling is hyperactivated in Lenvatinib-resistant HCC cells, C504244 was tested in combination with Lenvatinib and found to markedly sensitize these resistant cells to Lenvatinib treatment both and .
C504244 represents a promising agent that effectively inhibits β-catenin signaling, thereby impairing CSCs properties and reversing Lenvatinib resistance in HCC cells. These findings suggest that C504244 may serve as a potential therapeutic agent for HCC.
肝癌仍然是一项重大的全球健康负担，其中肝细胞癌（HCC）约占肝癌病例的80%。癌症干细胞（CSCs）在HCC的起始、进展、转移及对治疗的抗性中起关键作用，使其成为新型治疗干预的关键靶点。然而，特异性靶向HCC中CSCs的有效药物仍然有限。本研究的目的是鉴定和表征抑制CSCs特性并克服HCC耐药性的新型小分子。
功能试验评估了C504244对肿瘤球形成、癌细胞增殖和迁移的影响。对经C504244处理的HCC细胞进行RNA测序，以研究基因表达谱的变化。分析Wnt信号通路的下游靶点以确定通路抑制情况。进行免疫共沉淀（Co-IP）以评估C504244是否破坏HCC细胞中β-连环蛋白与转录因子4（TCF4）之间的相互作用。使用对乐伐替尼耐药的HCC细胞系评估C504244与乐伐替尼联合使用的疗效。
C504244显著抑制HCC细胞的肿瘤球形成、增殖和迁移。转录组分析显示，C504244处理导致Wnt信号通路受到显著抑制，下游靶基因表达相应下调。从机制上讲，C504244破坏了β-连环蛋白/TCF4复合物的形成，这可能导致转录活性降低。由于β-连环蛋白信号在对乐伐替尼耐药的HCC细胞中过度激活，因此对C504244与乐伐替尼联合使用进行了测试，发现其能使这些耐药细胞对乐伐替尼治疗显著敏感。
C504244是一种有前景的药物，可有效抑制β-连环蛋白信号，从而损害CSCs特性并逆转HCC细胞中的乐伐替尼耐药性。这些发现表明C504244可能作为HCC的潜在治疗药物。
