Tan Ying, Zhang Xian, Wang Jin, Xiao Xue, Yang Jin-Lin
Department of Gastroenterology and Hepatology, West China Hospital of Sichuan University, Chengdu, Sichuan, China.
Sichuan University-Oxford University Huaxi Gastrointestinal Cancer Center, West China Hospital of Sichuan University, Chengdu, Sichuan, China.
Dig Dis Sci. 2025 Sep 10. doi: 10.1007/s10620-025-09386-0.
Liver metastasis significantly contributes to poor survival in patients with colorectal cancer (CRC), posing therapeutic challenges due to limited understanding of its mechanisms. We aimed to identify a potential target critical for CRC liver metastasis.
We analyzed the Gene Expression Omnibus (GEO) and the Cancer Genome Atlas (TCGA) databases and identified EphrinA3 (EFNA3) as a potential clinically relevant target. The interacting proteins of EFNA3 were investigated by co-immunoprecipitation. The role of EphA3/EFNA3 axis were examined using EFNA3 knockdown and overexpressed CRC cells by transwell, wound healing, cell viability, colony formation and apoptosis assays, as well as subcutaneous and spleen injection tumour models in nude mice.
EFNA3 expression was significantly higher in CRC tissues than in para-carcinoma tissues, and elevated in metastatic tissues compared to primary CRC tissues. Higher EFNA3 was significantly correlated with advanced tumour stages and unfavourable clinical outcomes in CRC patients. Functional assays suggested that EFNA3 knockdown inhibited the migration, invasion and proliferation of CRC cells both in vitro and in vivo, while EFNA3 overexpression had opposite effects. Mechanistically, EphA3 was confirmed to bind to EFNA3, and EphA3/EFNA3 reverse signalling was found to promote epithelial-to-mesenchymal transition (EMT) by activating extracellular signal-regulated kinase (ERK) 1/2 signalling, thereby promoting metastasis.
Our findings suggest that EFNA3 promotes CRC metastasis, and that EphA3/EFNA3 signalling may promote EMT by activating the ERK signalling. These results indicate that the EphA3/EFNA3 axis could be a potential target for metastatic CRC.
肝转移显著导致结直肠癌（CRC）患者生存率低下，由于对其机制了解有限，给治疗带来了挑战。我们旨在确定一个对CRC肝转移至关重要的潜在靶点。
我们分析了基因表达综合数据库（GEO）和癌症基因组图谱（TCGA）数据库，并确定EphrinA3（EFNA3）为一个潜在的临床相关靶点。通过免疫共沉淀研究EFNA3的相互作用蛋白。使用EFNA3敲低和过表达的CRC细胞，通过Transwell实验、伤口愈合实验、细胞活力实验、集落形成实验和凋亡实验，以及裸鼠皮下和脾脏注射肿瘤模型，研究EphA3/EFNA3轴的作用。
EFNA3在CRC组织中的表达显著高于癌旁组织，且与原发性CRC组织相比，在转移组织中表达升高。较高的EFNA3与CRC患者的晚期肿瘤分期和不良临床结局显著相关。功能实验表明，EFNA3敲低在体外和体内均抑制CRC细胞的迁移、侵袭和增殖，而EFNA3过表达则有相反的作用。机制上，证实EphA3与EFNA3结合，并且发现EphA3/EFNA3反向信号通过激活细胞外信号调节激酶（ERK）1/2信号促进上皮-间质转化（EMT），从而促进转移。
我们的研究结果表明EFNA3促进CRC转移，并且EphA3/EFNA3信号可能通过激活ERK信号促进EMT。这些结果表明EphA3/EFNA3轴可能是转移性CRC的一个潜在靶点。
