Endosomal and lysosomal effects of desferrioxamine: protection of HeLa cells from hydrogen peroxide-induced DNA damage and induction of cell-cycle arrest.
The role of endosomal/lysosomal redox-active iron in H2O2-induced nuclear DNA damage as well as in cell proliferation was examined using the iron chelator desferrioxamine (DFO). Transient transfections of HeLa cells with vectors encoding dominant proteins involved in the regulation of various routes of endocytosis (dynamin and Rab5) were used to show that DFO (a potent and rather specific iron chelator) enters cells by fluid-phase endocytosis and exerts its effects by chelating redox-active iron present in the endosomal/lysosomal compartment. Endocytosed DFO effectively protected cells against H2O2-induced DNA damage, indicating the importance of endosomal/lysosomal redox-active iron in these processes. Moreover, exposure of cells to DFO in a range of concentrations (0.1 to 100 microM) inhibited cell proliferation in a fluid-phase endocytosis-dependent manner. Flow cytometric analysis of cells exposed to 100 microM DFO for 24 h showed that the cell cycle was transiently interrupted at the G2/M phase, while treatment for 48 h led to permanent cell arrest. Collectively, the above results clearly indicate that DFO has to be endocytosed by the fluid-phase pathway to protect cells against H2O2-induced DNA damage. Moreover, chelation of iron in the endosomal/lysosomal cell compartment leads to cell cycle interruption, indicating that all cellular labile iron is propagated through this compartment before its anabolic use is possible.
Doulias
Paschalis-Thomas
PT
Laboratory of Biological Chemistry, University of Ioannina Medical School, Ioannina, Greece.
Christoforidis
Savvas
S
Brunk
Ulf T
UT
Galaris
Dimitrios
D
D002453:Cell Cycle / Q000187:drug effects* / Q000502:physiology; D002455:Cell Division / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects* / Q000502:physiology; D003676:Deferoxamine / Q000494:pharmacology*; D034281:Dynamins / Q000235:genetics / Q000378:metabolism; D011992:Endosomes / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000009:adverse effects*; D007501:Iron / Q000378:metabolism; D007502:Iron Chelating Agents / Q000494:pharmacology; D000090463:Iron Deficiencies; D008247:Lysosomes / Q000187:drug effects*; D009154:Mutation; D013997:Time Factors; D020696:rab5 GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism
D007502:Iron Chelating Agents; D006861:Hydrogen Peroxide; D007501:Iron; D020696:rab5 GTP-Binding Proteins; D034281:Dynamins; D003676:Deferoxamine