Diazene JK-279 induces apoptosis-like cell death in human cervical carcinoma cells.
Diazene N-phenyl-2-(2-pyridinyl)diazenecarboxamide (JK-279) is a newly synthesized compound, cytotoxic for several tumor cell lines and their drug-resistant sublines. In human cervical carcinoma cells (HeLa), this compound reduced intracellular glutathione content and increased sensitivity to cisplatin. The aim of the present study was to elucidate the molecular mechanisms involved in the cytotoxic effect of diazene JK-279 on HeLa cells. Cytotoxicity was determined by the MTT method. Flow cytometry analysis showed that diazene JK-279 induces G(2)/M phase arrest, mediated by the increase in p21 expression, and accompanied by an alteration in the expression of survivin. The highest concentration of JK-279 altered nuclear morphology in intact cells, showing "apoptosis-like" features. No cleavage of procaspase-3, procaspase-9 and PARP, or altered expression of apoptotic proteins Bcl-2 and Bax were detected. At the same time, PS externalization and internucleosomal DNA cleavage were observed. Partial necrosis was detected as well. Our results demonstrate that cytotoxicity of diazene JK-279 is mostly the consequence of caspase-independent cell death, which is in some aspects "apoptosis-like". Taking into account the multiplicity of mechanisms used by cancer cells to prevent apoptosis, the drugs (like diazene JK-279) that would activate alternative cell death pathways could provide a useful tool for new types of cancer therapy.
Jakopec
S
S
Department of Molecular Biology, Rudjer Boskovic Institute, Bijenicka cesta 54, HR-10000 Zagreb, Croatia.
Dubravcic
K
K
Polanc
S
S
Kosmrlj
J
J
Osmak
M
M
D017304:Annexin A5 / Q000378:metabolism; D000970:Antineoplastic Agents / Q000633:toxicity*; D017209:Apoptosis; D001372:Aza Compounds / Q000633:toxicity*; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D016923:Cell Death; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002463:Cell Membrane Permeability; D002467:Cell Nucleus / Q000187:drug effects / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D024161:Collagen Type XI / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism; D011419:Propidium / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism; D011725:Pyridines / Q000633:toxicity*; D000077022:Survivin
D017304:Annexin A5; D000970:Antineoplastic Agents; D001372:Aza Compounds; C107489:BIRC5 protein, human; C499319:COL11A2 protein, human; D024161:Collagen Type XI; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; C509684:N-phenyl-2-(2-pyridinyl)diazenecarboxamide; D009363:Neoplasm Proteins; D011725:Pyridines; D000077022:Survivin; D011419:Propidium; C505407:CASP3 protein, human; C506433:CASP9 protein, human; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases; D016283:Proto-Oncogene Proteins p21(ras); D005978:Glutathione