Activation of the P2X7 receptor leads to a rapid, bidirectional flux of cations, causing broad range of biological responses including cytotoxicity. However, the mechanism of P2X7-mediated cytotoxicity remains largely unexplored. In our previous study, the lack of P2X7-mediated calcium response under normal conditions was found in P2X7(+) hematopoietic cell lines. In this study, the P2X7-mediated cytotoxicity in different type of cells (P2X7(-), P2X7(+) with calcium response, and P2X7(+) without calcium response) was investigated. Our results showed that P2X7 agonists, adenosine 5'-triphosphate (ATP) or 2',3'-O-(4 benzoylbenzoyl)-ATP, dose-dependently reduced the cell viability in all P2X7(+) cells tested, including J6-1, LCL, and Namalva cells which are negative for P2X7-mediated calcium response, although these effects were lower than those observed in KG1a cells which has normal P2X7 functions. The cytotoxic effect could be blocked by P2X7 antagonists, oxidized ATP and 1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine. In addition, externalization of phosphatidylserine could be detected in a time-dependent manner and apoptotic morphological changes could be observed after the activation of P2X7 receptor in J6-1 cells. Furthermore, P2X7-mediated pore formation could be detected in KG1a and J6-1 cells under low-ionic conditions, but not under low-divalent conditions. These effects could not be observed in P2X7(-) Ramos cells. These results suggested that P2X7 receptor-mediated cytotoxic effects may occur independent of calcium response.