Giannuzzo Andrea, Saccomano Mara, Napp Joanna, Ellegaard Maria, Alves Frauke, Novak Ivana
Section for Cell Biology and Physiology, August Krogh Building, Department of Biology, University of Copenhagen, Denmark.
Department of Molecular Biology of Neuronal Signals, Max Planck Institute for Experimental Medicine, Hermann-Rein-Straße 3, Göttingen, D-37075, Germany.
Int J Cancer. 2016 Dec 1;139(11):2540-52. doi: 10.1002/ijc.30380. Epub 2016 Aug 29.
The ATP-gated receptor P2X7 (P2X7R) is involved in regulation of cell survival and has been of interest in cancer field. Pancreatic ductal adenocarcinoma (PDAC) is a deadly cancer and new markers and therapeutic targets are needed. PDAC is characterized by a complex tumour microenvironment, which includes cancer and pancreatic stellate cells (PSCs), and potentially high nucleotide/side turnover. Our aim was to determine P2X7R expression and function in human pancreatic cancer cells in vitro as well as to perform in vivo efficacy study applying P2X7R inhibitor in an orthotopic xenograft mouse model of PDAC. In the in vitro studies we show that human PDAC cells with luciferase gene (PancTu-1 Luc cells) express high levels of P2X7R protein. Allosteric P2X7R antagonist AZ10606120 inhibited cell proliferation in basal conditions, indicating that P2X7R was tonically active. Extracellular ATP and BzATP, to which the P2X7R is more sensitive, further affected cell survival and confirmed complex functionality of P2X7R. PancTu-1 Luc migration and invasion was reduced by AZ10606120, and it was stimulated by PSCs, but not by PSCs from P2X7(-/-) animals. PancTu-1 Luc cells were orthotopically transplanted into nude mice and tumour growth was followed noninvasively by bioluminescence imaging. AZ10606120-treated mice showed reduced bioluminescence compared to saline-treated mice. Immunohistochemical analysis confirmed P2X7R expression in cancer and PSC cells, and in metaplastic/neoplastic acinar and duct structures. PSCs number/activity and collagen deposition was reduced in AZ10606120-treated tumours.
三磷酸腺苷（ATP）门控受体P2X7（P2X7R）参与细胞存活的调节，在癌症领域备受关注。胰腺导管腺癌（PDAC）是一种致命的癌症，需要新的标志物和治疗靶点。PDAC的特征是肿瘤微环境复杂，其中包括癌细胞和胰腺星状细胞（PSC），并且可能存在高核苷酸/底物周转率。我们的目的是在体外确定P2X7R在人胰腺癌细胞中的表达和功能，并在PDAC原位异种移植小鼠模型中应用P2X7R抑制剂进行体内疗效研究。在体外研究中，我们发现带有荧光素酶基因的人PDAC细胞（PancTu-1 Luc细胞）表达高水平的P2X7R蛋白。变构P2X7R拮抗剂AZ10606120在基础条件下抑制细胞增殖，表明P2X7R具有持续性活性。细胞外ATP和对P2X7R更敏感的BzATP进一步影响细胞存活，并证实了P2X7R的复杂功能。AZ10606120可降低PancTu-1 Luc细胞的迁移和侵袭能力，PSC可刺激其迁移和侵袭，但P2X7基因敲除（P2X7(-/-)）动物的PSC则无此作用。将PancTu-1 Luc细胞原位移植到裸鼠体内，通过生物发光成像对肿瘤生长进行无创监测。与生理盐水处理的小鼠相比，AZ10606120处理的小鼠生物发光减弱。免疫组织化学分析证实P2X7R在癌细胞、PSC细胞以及化生/肿瘤性腺泡和导管结构中均有表达。AZ10606120处理的肿瘤中PSC数量/活性和胶原沉积减少。
