Nuñez-Ríos José David, Reyna-Jeldes Mauricio, Mata-Martínez Esperanza, Campos-Contreras Anaí Del Rocío, Lazcano-Sánchez Iván, González-Gallardo Adriana, Díaz-Muñoz Mauricio, Coddou Claudio, Vázquez-Cuevas Francisco G
Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Querétaro, México.
Departamento de Ciencias Biomédicas, Facultad de Medicina, Universidad Católica del Norte, Coquimbo, Chile.
PLoS One. 2024 Jun 13;19(6):e0304062. doi: 10.1371/journal.pone.0304062. eCollection 2024.
ATP is actively maintained at high concentrations in cancerous tissues, where it promotes a malignant phenotype through P2 receptors. In this study, we first evaluated the effect of extracellular ATP depletion with apyrase in SKOV-3, a cell line derived from metastatic ovarian carcinoma. We observed a decrease in cell migration and an increase in transepithelial electrical resistance and cell markers, suggesting a role in maintaining a mesenchymal phenotype. To identify the P2 receptor that mediated the effects of ATP, we compared the transcript levels of some P2 receptors and found that P2RX7 is three-fold higher in SKOV-3 cells than in a healthy cell line, namely HOSE6-3 (from human ovarian surface epithelium). Through bioinformatic analysis, we identified a higher expression of the P2RX7 transcript in metastatic tissues than in primary tumors; thus, P2X7 seems to be a promising effector for the malignant phenotype. Subsequently, we demonstrated the presence and functionality of the P2X7 receptor in SKOV-3 cells and showed through pharmacological approaches that its activity promotes cell migration and contributes to maintaining a mesenchymal phenotype. P2X7 activation using BzATP increased cell migration and abolished E-cadherin expression. On the other hand, a series of P2X7 receptor antagonists (A438079, BBG and OxATP) decreased cell migration. We used a CRISPR-based knock-out system directed to P2RX7. According to the results of our wound-healing assay, SKOV3-P2X7KO cells lacked receptor-mediated calcium mobilization and decreased migration. Altogether, these data let us propose that P2X7 receptor is a regulator for cancer cell migration and thus a potential drug target.
三磷酸腺苷（ATP）在癌组织中被积极维持在高浓度水平，在那里它通过P2受体促进恶性表型。在本研究中，我们首先评估了用腺苷三磷酸双磷酸酶消耗细胞外ATP对SKOV-3细胞系的影响，SKOV-3是一种源自转移性卵巢癌的细胞系。我们观察到细胞迁移减少，跨上皮电阻和细胞标志物增加，这表明其在维持间充质表型中发挥作用。为了确定介导ATP作用的P2受体，我们比较了一些P2受体的转录水平，发现P2RX7在SKOV-3细胞中的表达比在健康细胞系即HOSE6-3（源自人卵巢表面上皮）中高3倍。通过生物信息学分析，我们发现在转移组织中P2RX7转录本的表达高于原发性肿瘤；因此，P2X7似乎是恶性表型的一个有前景的效应因子。随后，我们证明了SKOV-3细胞中P2X7受体的存在和功能，并通过药理学方法表明其活性促进细胞迁移并有助于维持间充质表型。使用2'-（3-苯甲酰苯甲酰）-ATP（BzATP）激活P2X7可增加细胞迁移并消除E-钙黏蛋白的表达。另一方面，一系列P2X7受体拮抗剂（A438079、BBG和氧化三磷酸腺苷（OxATP））可减少细胞迁移。我们使用了针对P2RX7的基于CRISPR的基因敲除系统。根据我们的伤口愈合试验结果，SKOV3-P2X7基因敲除细胞缺乏受体介导的钙动员且迁移减少。总之，这些数据使我们提出P2X7受体是癌细胞迁移的调节因子，因此是一个潜在的药物靶点。
