Stem-like cells (CSCs) have a tumour-initiating capacity and play critical role in tumour metastasis, relapse and resistance to therapy. The ectoenzyme CD73, encoded by the NT5E gene, which catalyses the hydrolysis of AMP into adenosine, has been associated to an immunosuppressive tumour microenvironment, tumour cell adhesion and migration. Therefore, we investigated the expression and activity of CD73 in sphere-forming cells from cervical cancer in comparison to monolayer cells in vitro. In addition, in silico analysis was performed to determine the expression of CD73 and other members of purinergic signalling in CSC-like population derived from different tumour types in comparison to monolayer cells. CD73 protein expression levels and functionality in SiHa cells were analysed by flow cytometry and enzymatic assay, respectively. In silico investigation was performed through the analysis of seven datasets from different tumour types using GEO database. In vitro analysis showed a decreased CD73 protein expression and enzymatic activity in cervical spheres, when compared to monolayers. In addition, when sphere-derived cells are re-plated as monolayer culture, the CD73 expression and activity are restored. Supporting the in vitro results, in silico analysis showed that three-dimensional spheres derived from cervical, thyroid and breast cancer presented decreased expression of CD73, when compared to their adherent counterparts. The decreased expression of CD73 in sphere-derived cells or CSC-enriched population reinforce its important role in cell adhesion, tumour spreading ability and metastasis, suggesting CD73 as potential target to be further investigated in cervical cancer.

中文翻译：

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三维微环境改变宫颈癌中 CD73 的表达

干细胞样细胞 (CSC) 具有启动肿瘤的能力，并在肿瘤转移、复发和治疗抵抗中起关键作用。胞外酶 CD73，由NT5E编码催化AMP水解为腺苷的基因与免疫抑制性肿瘤微环境、肿瘤细胞粘附和迁移有关。因此，与体外单层细胞相比，我们研究了 CD73 在来自宫颈癌的球体形成细胞中的表达和活性。此外，与单层细胞相比，进行了计算机分析以确定 CD73 和其他嘌呤能信号成员在源自不同肿瘤类型的 CSC 样群体中的表达。分别通过流式细胞术和酶法分析了 SiHa 细胞中 CD73 蛋白的表达水平和功能。通过使用 GEO 数据库分析来自不同肿瘤类型的七个数据集，进行了计算机模拟研究。体外分析表明，与单层相比，宫颈球中 CD73 蛋白表达和酶活性降低。此外，当球体衍生细胞作为单层培养物重新铺板时，CD73 的表达和活性得以恢复。支持体外结果，计算机分析表明，与粘附的对应物相比，来自宫颈癌、甲状腺癌和乳腺癌的三维球体的 CD73 表达降低。CD73 在球体衍生细胞或富含 CSC 的群体中表达的降低加强了其在细胞粘附、肿瘤扩散能力和转移中的重要作用，表明 CD73 是宫颈癌中需要进一步研究的潜在靶点。当球体衍生细胞作为单层培养物重新铺板时，CD73 表达和活性得以恢复。支持体外结果，计算机分析表明，与粘附的对应物相比，来自宫颈癌、甲状腺癌和乳腺癌的三维球体的 CD73 表达降低。CD73 在球体衍生细胞或富含 CSC 的群体中表达的降低加强了其在细胞粘附、肿瘤扩散能力和转移中的重要作用，表明 CD73 是宫颈癌中需要进一步研究的潜在靶点。当球体衍生细胞作为单层培养物重新铺板时，CD73 表达和活性得以恢复。支持体外结果，计算机分析表明，与粘附的对应物相比，来自宫颈癌、甲状腺癌和乳腺癌的三维球体的 CD73 表达降低。CD73 在球体衍生细胞或富含 CSC 的群体中表达的降低加强了其在细胞粘附、肿瘤扩散能力和转移中的重要作用，表明 CD73 是宫颈癌中需要进一步研究的潜在靶点。与他们的追随者相比。CD73 在球体衍生细胞或富含 CSC 的群体中表达的降低加强了其在细胞粘附、肿瘤扩散能力和转移中的重要作用，表明 CD73 是宫颈癌中需要进一步研究的潜在靶点。与他们的追随者相比。CD73 在球体衍生细胞或富含 CSC 的群体中表达的降低加强了其在细胞粘附、肿瘤扩散能力和转移中的重要作用，表明 CD73 是宫颈癌中需要进一步研究的潜在靶点。