z-val-ala-asp(ome)-fluoromethylketone and Necrosis

Apoptosis is triggered by the activation of caspases and characterized by chromatin condensation and nuclear fragmentation (type II nuclear morphology). Necrosis is depicted by a gain in cell volume (oncosis), swelling of organelles, plasma membrane leakage, and subsequent loss of intracellular contents. Although considered as different cell death entities, there is an overlap between apoptosis and necrosis. In this sense, mounting evidence suggests that both processes can be morphological expressions of a common biochemical network known as "apoptosis-necrosis continuum." To gain insight into the events driving the apoptosis-necrosis continuum, apoptotically proficient cells were screened facing several apoptotic inducers for the absence of type II apoptotic nuclear morphologies. Chelerythrine was selected for further studies based on its cytotoxicity and the lack of apoptotic nuclear alterations. Chelerythrine triggered an early plasma membrane leakage without condensed chromatin aggregates. Ultrastructural analysis revealed that chelerythrine-mediated cytotoxicity was compatible with a necrotic-like type of cell death. Biochemically, chelerythrine induced the activation of caspases. Moreover, the inhibition of caspases prevented chelerythrine-triggered necrotic-like cell death. Compared with staurosporine, chelerythrine induced stronger caspase activation detectable at earlier times. After using a battery of chemicals, we found that high concentrations of thiolic antioxidants fully prevented chelerythrine-driven caspase activation and necrotic-like cell death. Lower amounts of thiolic antioxidants partially prevented chelerythrine-mediated cytotoxicity and allowed cells to display type II apoptotic nuclear morphology correlating with a delay in caspase-3 activation. Altogether, these data support that an early and pronounced activation of caspases can drive cells to undergo a form of necrotic-like regulated cell death.

Topics: Amino Acid Chloromethyl Ketones; Antibodies, Monoclonal; Antineoplastic Agents; Apoptosis; Benzophenanthridines; Carrier Proteins; Caspases; Cell Line, Tumor; Chromatin; Colchicine; Enzyme Activation; Enzyme Inhibitors; Gene Expression Regulation; Humans; Microfilament Proteins; Necrosis; Neurons; Nocodazole; Peptidomimetics; Quinolines; Rotenone; Signal Transduction; Staurosporine; Thapsigargin

Topics: Amino Acid Chloromethyl Ketones; Apoptosis; Caspase Inhibitors; Caspases; Cathepsin B; Cell Survival; Dipeptidases; Dipeptides; Dose-Response Relationship, Drug; Enzyme Inhibitors; HL-60 Cells; Humans; Intracellular Membranes; Lysosomes; Necrosis; Signal Transduction; Staurosporine