RESUMO
Evasion of DNA damage-induced cell death, via mutation of the p53 tumor suppressor or overexpression of prosurvival Bcl-2 family proteins, is a key step toward malignant transformation and therapeutic resistance. We report that depletion or acute inhibition of checkpoint kinase 1 (Chk1) is sufficient to restore gamma-radiation-induced apoptosis in p53 mutant zebrafish embryos. Surprisingly, caspase-3 is not activated prior to DNA fragmentation, in contrast to classical intrinsic or extrinsic apoptosis. Rather, an alternative apoptotic program is engaged that cell autonomously requires atm (ataxia telangiectasia mutated), atr (ATM and Rad3-related) and caspase-2, and is not affected by p53 loss or overexpression of bcl-2/xl. Similarly, Chk1 inhibitor-treated human tumor cells hyperactivate ATM, ATR, and caspase-2 after gamma-radiation and trigger a caspase-2-dependent apoptotic program that bypasses p53 deficiency and excess Bcl-2. The evolutionarily conserved "Chk1-suppressed" pathway defines a novel apoptotic process, whose responsiveness to Chk1 inhibitors and insensitivity to p53 and BCL2 alterations have important implications for cancer therapy.
Assuntos
Apoptose , Caspase 2/metabolismo , Dano ao DNA , Proteínas Quinases/metabolismo , Transdução de Sinais , Peixe-Zebra/metabolismo , Animais , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Caspase 3/metabolismo , Linhagem Celular Tumoral , Quinase 1 do Ponto de Checagem , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/metabolismo , Embrião não Mamífero/efeitos da radiação , Inibidores Enzimáticos/farmacologia , Raios gama , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismoAssuntos
Praias , Exantema/diagnóstico , Exantema/etiologia , Infecções por Uncinaria/complicações , Albendazol/uso terapêutico , Ancylostoma/patogenicidade , Animais , Antiparasitários/uso terapêutico , Antiprotozoários/uso terapêutico , Feminino , Humanos , Ivermectina/uso terapêutico , Pessoa de Meia-Idade , Prurido/etiologia , ViagemRESUMO
PURPOSE: The present study explored the causal relationship between stressor exposure/stress neuropeptide activation and avoidant exploratory phenotype/enhanced seizure susceptibility in an animal model of epilepsy. METHODS: The olfactory detection and investigation phenotype of seizure susceptible El (epilepsy) strain and nonsusceptible ddY control mice was first evaluated in untreated mice. In a second series of experiments, the olfactory exploration phenotype, food intake/body weight regulation, circadian locomotor activity, and seizure susceptibility were assessed over a 14-day period following central administration of the neurotoxin saporin alone or a conjugate of the stress neuropeptide, corticotropin releasing factor (CRF), and saporin (CRF-SAP) which impairs CRF system function following central administration. RESULTS: In support of the main experimental hypothesis, administration of CRF-SAP in El mice reduced handling-induced seizure susceptibility by 75% for up to 2 weeks following treatment. Similarly, El mice were slow to detect a cache of buried food pellets relative to ddY controls and this exploratory deficit was reversed 3 days following administration of CRF-SAP. Efficacy of CRF-SAP treatment was confirmed using CRF immunohistochemistry, which revealed suppression of brain CRF content in El mice treated with CRF-SAP relative to El controls. Other functional and persistent effects of CRF-SAP included increased locomotor activity and hyperphagia. CONCLUSIONS: Taken together, these results support strongly the possibility that activated brain stress neuropeptide systems are necessary for the expression of motivational and neurological perturbations in seizure susceptible El mice.