Department of Medical Sciences, University of Ferrara, Ferrara, Italy.
Methods Mol Biol. 2022;2510:303-314. doi: 10.1007/978-1-0716-2384-8_17.
The tumor microenvironment is rich in components that strongly influence cancer cell survival. One of the pivotal molecules present at the tumor bed is ATP, which has an essential role in promoting cancer proliferation and metastasis and immune responses via its receptor P2X7. Several studies have proved the efficacy of P2X7 pharmacological blockade in inhibiting primary and metastatic tumor growth in preclinical models. Here we describe the experimental procedures that we optimized to test P2X7 roles in carcinogenesis by antagonist administration. Special attention is paid to their concentrations and routes of administration. The depicted in vitro models include cell count and viability assays, which are useful to test P2X7 roles in cell proliferation and vitality, and the soft agar colony formation test that allows investigation of the transforming and invading abilities of tumor cells. We also describe systemic and intramass administration of P2X7 blockers in murine models of melanoma and leukemia. Both xenotransplant and syngeneic experimental tumor models are detailed.
肿瘤微环境富含强烈影响癌细胞存活的成分。肿瘤床存在的关键分子之一是 ATP,它通过其受体 P2X7 在促进癌症增殖、转移和免疫反应中发挥重要作用。多项研究已经证明了 P2X7 药理学阻断在抑制临床前模型中原发性和转移性肿瘤生长方面的功效。在这里,我们描述了我们优化的实验程序,以通过拮抗剂给药来测试 P2X7 在致癌作用中的作用。特别注意它们的浓度和给药途径。所描述的体外模型包括细胞计数和活力测定,这些测定可用于测试 P2X7 在细胞增殖和活力中的作用,以及软琼脂集落形成试验,该试验可研究肿瘤细胞的转化和侵袭能力。我们还描述了 P2X7 阻滞剂在黑色素瘤和白血病的小鼠模型中的系统和瘤内给药。详细介绍了异种移植和同基因实验肿瘤模型。