ROS generate dalle NAD(P)H ossidasi nella segnalazione redox in linee cellulari leucemiche

Discovery of the Nox family has led to the concept that ROS are “intentionally” generated and are biologically functional in various cell types. Over the last decades, ROS have been shown to be involved in several physiological and pathological processes and ROS producing enzymes have been suggested as a target for drug development. The mechanism involved in the prosurvival effect of cytokines on the human acute myeloid leukaemia cell lines M07e and B1647 is investigated. A decrease in intracellular reactive oxygen species (ROS) content, glucose transport activity and cell survival was observed in the presence of inhibitors of plasma membrane ROS sources, such as DPI and apocynin, and by small interference RNA for NOX2 in M07e cells. Furthermore, Nox generated ROS are required to sustain B1647 cell viability and proliferation; in fact, antioxidants such as EUK-134 or Nox inhibitors and siRNA direct cells to apoptotic cell death, suggesting that manipulation of cellular NOX2 and NOX4 could affect survival of leukemic cells. Moreover, hydrogen peroxide has been long thought to be freely diffusible but recent evidence suggest that specific mammalian aquaporin homologues (AQP8) possess the capacity to channel H2O2 across membrane. In this thesis is shown that inhibition of aquaporins diminishes intracellular ROS accumulation either when H2O2 is produced by Nox enzymes or when is added exogenously to the medium. These data suggest that specific inhibition of Nox enzymes and AQP8 could be an interesting novel anti-leukemic strategy.