细胞外三磷酸腺苷 (ATP) 传导广泛的细胞信号传导的复杂动态系统。外切核苷酸酶是具有核苷酸水解能力的酶，可在生理和病理条件下调节 ATP 水平，从而在所谓的嘌呤能信号传导中发挥关键作用。据报道，癌症中外切核苷酸酶的表达发生了改变，而外切核苷酸三磷酸二磷酸水解酶 (NTPDase) 家族及其最著名的形式 NTPDase1 (CD39) 是癌症免疫治疗的目标。 CD39-CD73 酶的串联负责在肿瘤微环境中产生免疫抑制腺苷，抑制策略引起了人们的极大兴趣。类器官已成为研究肿瘤的非常方便的模型，因为它们是三维培养物，保留了组织的许多特征。本研究旨在有助于改进健康和疾病条件下核酸酶研究所需的方法和分子工具。这项研究是在子宫内膜癌细胞模型中进行的，可以扩展到涉及嘌呤能系统的其他类型的肿瘤和病理学。我们从过表达 NTPDase2 (CD39L1) 和 NTPDase3 (CD39L3) 的子宫内膜癌细胞中生成了类器官，作为与 EGFP 的融合蛋白，并通过采用原位细胞化学方案进行了功能测定。这使我们能够同时检测酶活性和蛋白质表达，并证明类器官可用于测试核酸外切酶抑制剂，这一结果可用于开发新的癌症治疗方案。©作者 2024。开放获取。本文根据知识共享 CC-BY 国际许可证获得许可。

Extracellular adenosine triphosphate (ATP) conducts a complex dynamic system of broadly represented cell signaling. Ectonucleotidases are the enzymes with nucleotide hydrolytic ability that regulate ATP levels in physiological and pathological conditions, thus playing a key role in the so-called purinergic signaling. Altered ectonucleotidase expression has been reported in cancer, and the ectonucleoside triphosphate diphosphohydrolase (NTPDase) family of enzymes, with its best-known form NTPDase1 (CD39), is targeted in cancer immunotherapy. The tandem of enzymes CD39-CD73 is responsible for the generation of immunosuppressive adenosine in the tumor microenvironment, and inhibition strategies are of great interest. Organoids have emerged as very convenient models for the study of tumors since they are three-dimensional cultures that retain many of the features of tissue. The present study aims to contribute to improving the methodology and the molecular tools needed for the study of ectonucleotidases in healthy and disease conditions. The study, performed in an endometrial cancer cell model, could be extended to other types of tumors and pathologies in which the purinergic system is involved. We generated organoids from endometrial cancer cells overexpressing NTPDase2 (CD39L1) and NTPDase3 (CD39L3) as fusion proteins with EGFP, and we performed functional assays by adapting in situ cytochemistry protocols. This allowed us to simultaneously detect enzyme activity and protein expression and to demonstrate that organoids can be used to test ectonucleotidase inhibitors-a result that can be used to develop new cancer treatment options.©The Author(s) 2024. Open Access. This article is licensed under a Creative Commons CC-BY International License.