PDF file, 72KB, Fig. S4. Effect of P2X7 silencing on ACN neuroblastoma growth. Panel A, P2X7 expression in ACN neuroblastoma cells transfected with P2X7-specific or scrambled shRNA. Cell protein was extracted, run on SDS PAGE, blotted and stained with an anti-P2X7-selective polyclonal Ab. Panel B, densitometric analysis of P2X7 bands. Panel C, volumes of tumors excised at d 18 post-inoculum.
ARTICLE ABSTRACT
The P2X7 receptor is an ATP-gated ion channel known for its cytotoxic activity. However, recent evidence suggests a role for P2X7 in cell proliferation. Here, we found that P2X7 exhibits significant growth-promoting effects in vivo. Human embryonic kidney cells expressing P2X7 exhibited a more tumorigenic and anaplastic phenotype than control cells in vivo, and the growth rate and size of these tumors were significantly reduced by intratumoral injection of the P2X7 inhibitor–oxidized ATP. The accelerated growth of P2X7-expressing tumors was characterized by increased proliferation, reduced apoptosis, and a high level of activated transcription factor NFATc1. These tumors also showed a more developed vascular network than control tumors and secreted elevated amounts of VEGF. The growth and neoangiogenesis of P2X7-expressing tumors was blocked by intratumoral injection of the VEGF-blocking antibody Avastin (bevacizumab), pharmacologic P2X7 blockade, or P2X7 silencing in vivo. Immunohistochemistry revealed strong P2X7 positivity in several human cancers. Together, our findings provide direct evidence that P2X7 promotes tumor growth in vivo. Cancer Res; 72(12); 2957–69. ©2012 AACR.
