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Figure S1 from Accelerated Tumor Progression in Mice Lacking the ATP Receptor P2X7

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posted on 2023-03-30, 23:31 authored by Elena Adinolfi, Marina Capece, Alessia Franceschini, Simonetta Falzoni, Anna L. Giuliani, Alessandra Rotondo, Alba C. Sarti, Massimo Bonora, Susanne Syberg, Domenica Corigliano, Paolo Pinton, Niklas R. Jorgensen, Luigi Abelli, Laura Emionite, Lizzia Raffaghello, Vito Pistoia, Francesco Di Virgilio

Figure S1. Characterization of P2X7R shRNA B16 cells. A, densitometry of P2X7R protein expression. B, representative Western blot. C, Fura-2 measurement of Benzoyl ATP (BzATP)-triggered intracellular Ca2+ increase. D, Averages intracellular Ca2+ increases in wt and P2X7R-silenced B16 cells. Data are averages + SEM, n = 5. ***, p < 0.001. E, rate of proliferation of wt and P2X7R-silenced B16 cell cultures. Intracellular Ca2+ changes and proliferation rates were measured as described in Adinolfi et al., 2005, Mol Biol Cell 16:3260-3272.

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ARTICLE ABSTRACT

The ATP receptor P2X7 (P2X7R or P2RX7) has a key role in inflammation and immunity, but its possible roles in cancer are not firmly established. In the present study, we investigated the effect of host genetic deletion of P2X7R in the mouse on the growth of B16 melanoma or CT26 colon carcinoma cells. Tumor size and metastatic dissemination were assessed by in vivo calliper and luciferase luminescence emission measurements along with postmortem examination. In P2X7R-deficient mice, tumor growth and metastatic spreading were accelerated strongly, compared with wild-type (wt) mice. Intratumoral IL-1β and VEGF release were drastically reduced, and inflammatory cell infiltration was abrogated nearly completely. Similarly, tumor growth was also greatly accelerated in wt chimeric mice implanted with P2X7R-deficient bone marrow cells, defining hematopoietic cells as a sufficient site of P2X7R action. Finally, dendritic cells from P2X7R-deficient mice were unresponsive to stimulation with tumor cells, and chemotaxis of P2X7R-less cells was impaired. Overall, our results showed that host P2X7R expression was critical to support an antitumor immune response, and to restrict tumor growth and metastatic diffusion. Cancer Res; 75(4); 635–44. ©2014 AACR.