title: The DNA methyltransferase inhibitors zebularine and decitabine induce mitochondria-mediated apoptosis and DNA damage in p53 mutant leukemic T cells.
abstract: DNA methyltransferase (DNMT)-inhibiting nucleoside analogs reactivate the expression of tumor suppressor genes and apoptosis-related genes silenced by methylation, thus favoring the induction of apoptosis in tumor cells. Moreover, induction of DNA damage seems to contribute to their antitumor effect. However, the apoptotic signaling pathway induced by these demethylating drugs is not well understood. Here, we have investigated the induction of apoptosis by two nucleoside DNMT inhibitors, decitabine and zebularine, in leukemic T cells. Both inhibitors induced caspase-dependent apoptosis in Jurkat, CEM-6 and MOLT-4 leukemia T cell lines, all with mutant p53, whereas resting and activated normal T lymphocytes were highly resistant to these demethylating agents. Although decitabine and zebularine showed different ability to induce apoptosis and cell cycle arrest among the three cell lines, they similarly activated the intrinsic apoptotic pathway by inducing mitochondrial alterations such as Bak activation, loss of transmembrane potential and generation of reactive oxygen species (ROS). Accordingly, Bcl-2- and Bcl-x(L) -overexpressing Jurkat cells, as well as caspase-9-deficient Jurkat cells, were resistant to apoptosis induced by decitabine and zebularine. Interestingly, ROS production seemed to be necessary for the induction of apoptosis. Apoptotic events, such as Bak and caspase activation, started as soon as 20 hr after treatment with either decitabine or zebularine. In addition, progression of apoptosis triggered by both DNMT inhibitors was paralleled by the induction of DNA damage. Our results suggest that the mitochondrial apoptotic pathway activated by decitabine and zebularine in p53 mutant leukemic T cells depends mainly on the induction of DNA damage.
authors: Unidad de Inmunología, IBIMER, Universidad de Granada, Granada, Spain.
authors: María J
authors: MJ
authors: Ruiz-Magaña
authors: Jose M
authors: JM
authors: Rodríguez-Vargas
authors: Jorge C
authors: JC
authors: Morales
authors: Manuel A
authors: MA
authors: Saldivia
authors: Klaus
authors: K
authors: Schulze-Osthoff
authors: Carmen
authors: C
authors: Ruiz-Ruiz
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D053453:Caspase 9 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D003562:Cytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000037:antagonists & inhibitors*; D004249:DNA Damage / Q000187:drug effects*; D000077209:Decitabine; D016158:Genes, p53; D006801:Humans; D015458:Leukemia, T-Cell / Q000235:genetics* / Q000473:pathology; D008928:Mitochondria / Q000502:physiology*; D009154:Mutation; D017382:Reactive Oxygen Species / Q000378:metabolism; D013601:T-Lymphocytes / Q000473:pathology
chemical_list: D017382:Reactive Oxygen Species; D003562:Cytidine; D000077209:Decitabine; C009131:pyrimidin-2-one beta-ribofuranoside; D004248:DNA (Cytosine-5-)-Methyltransferases; D053453:Caspase 9; D001374:Azacitidine