Background/Objectives: Hypoxia promotes esophageal adenocarcinoma (EAC) aggressiveness through stabilization of hypoxia-inducible factor-1α (HIF-1α), which regulates pro-survival, pro-metastatic, and immunosuppressive pathways, including the ectoenzyme CD73 (NT5E). Although CD73 is a known hypoxia-responsive gene, its functional integration with HIF-1α signaling in EAC remains incompletely understood. This study aimed to define the relationship between HIF-1α and CD73 in EAC and to evaluate the therapeutic potential of their combined inhibition. Methods: Gene expression and survival analyses were performed using CCLE and TCGA-ESCA datasets. CD73 and HIF-1α expression were evaluated in EAC patient tissues by immunohistochemistry. EAC cell lines were subjected to hypoxic conditions with genetic or pharmacologic inhibition of HIF-1α and/or CD73. Cell viability, migration, angiogenesis, VEGF secretion, and purinergic metabolite levels were assessed using luminescence assays, Boyden chamber migration assays, endothelial tube formation assays, ELISA, and targeted LC-MS/MS, respectively. Results: NT5E expression was transcriptionally upregulated by HIF-1α under hypoxia and correlated with advanced disease stage and poor overall survival in EAC patients. While CD73 inhibition alone modestly reduced EAC cell viability, combined inhibition of HIF-1α and CD73 synergistically decreased tumor cell survival, particularly under hypoxic conditions, and significantly altered extracellular adenosine metabolism. Dual targeting further suppressed migration, reduced VEGF secretion, and impaired angiogenic signaling, indicating disruption of tumor microenvironmental pathways critical for metastasis and immune evasion. Conclusions: These findings identify CD73 as a direct hypoxia-responsive effector of HIF-1α in EAC and demonstrate that dual inhibition of HIF-1α and CD73 synergistically disrupts tumor cell survival and pro-metastatic signaling. This combinatorial strategy represents a mechanistically integrated therapeutic approach to overcome hypoxia-driven resistance in esophageal adenocarcinoma.
背景/目的：缺氧通过稳定缺氧诱导因子-1α（HIF-1α）促进食管腺癌（EAC）的侵袭性，该因子调控促生存、促转移和免疫抑制通路，包括外切酶CD73（NT5E）。尽管CD73是已知的缺氧响应基因，但其在EAC中与HIF-1α信号通路的功能整合机制尚未完全阐明。本研究旨在明确HIF-1α与CD73在EAC中的关系，并评估联合抑制两者的治疗潜力。方法：利用CCLE和TCGA-ESCA数据集进行基因表达与生存分析。通过免疫组化检测EAC患者组织中CD73和HIF-1α的表达。对EAC细胞系进行缺氧处理，并采用基因或药物手段抑制HIF-1α和/或CD73。分别通过发光检测、Boyden小室迁移实验、内皮细胞管形成实验、ELISA及靶向LC-MS/MS技术评估细胞活力、迁移能力、血管生成、VEGF分泌及嘌呤代谢物水平。结果：缺氧条件下HIF-1α转录上调NT5E表达，且其表达与EAC患者疾病晚期和总生存期缩短相关。单独抑制CD73仅适度降低EAC细胞活力，而联合抑制HIF-1α与CD73可协同降低肿瘤细胞存活率（尤其在缺氧条件下），并显著改变细胞外腺苷代谢。双重靶向抑制进一步削弱细胞迁移能力、减少VEGF分泌、破坏血管生成信号，表明其对肿瘤微环境中转移和免疫逃逸关键通路具有阻断作用。结论：本研究证实CD73是EAC中HIF-1α的直接缺氧响应效应因子，并证明联合抑制HIF-1α与CD73能协同破坏肿瘤细胞存活及促转移信号传导。该联合策略为克服食管腺癌中缺氧驱动性耐药提供了机制整合型治疗新途径。
肿瘤(癌症)患者之家