Inhibition of autophagy enhances DNA damage-induced apoptosis by disrupting CHK1-dependent S phase arrest.

Liou, Jong-Shian; Wu, Yi-Chen; Yen, Wen-Yen; Tang, Yu-Shuan; Kakadiya, Rajesh B; Su, Tsann-Long; Yih, Ling-Huei

Liou, Jong-Shian; Wu, Yi-Chen; Yen, Wen-Yen; Tang, Yu-Shuan; Kakadiya, Rajesh B; Su, Tsann-Long; Yih, Ling-Huei.

Afiliación

Liou JS; Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan, ROC.
Wu YC; Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan, ROC.
Yen WY; Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan, ROC.
Tang YS; Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan, ROC.
Kakadiya RB; Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan, ROC.
Su TL; Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan, ROC.
Yih LH; Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan, ROC. Electronic address: lhyih@gate.sinica.edu.tw.

Toxicol Appl Pharmacol ; 278(3): 249-58, 2014 Aug 01.

Article en En | MEDLINE | ID: mdl-24823293

RESUMEN

DNA damage has been shown to induce autophagy, but the role of autophagy in the DNA damage response and cell fate is not fully understood. BO-1012, a bifunctional alkylating derivative of 3a-aza-cyclopenta[a]indene, is a potent DNA interstrand cross-linking agent with anticancer activity. In this study, BO-1012 was found to reduce DNA synthesis, inhibit S phase progression, and induce phosphorylation of histone H2AX on serine 139 (Î³H2AX) exclusively in S phase cells. Both CHK1 and CHK2 were phosphorylated in response to BO-1012 treatment, but only depletion of CHK1, but not CHK2, impaired BO-1012-induced S phase arrest and facilitated the entry of Î³H2AX-positive cells into G2 phase. CHK1 depletion also significantly enhanced BO-1012-induced cell death and apoptosis. These results indicate that BO-1012-induced S phase arrest is a CHK1-dependent pro-survival response. BO-1012 also resulted in marked induction of acidic vesicular organelle (AVO) formation and microtubule-associated protein 1 light chain 3 (LC3) processing and redistribution, features characteristic of autophagy. Depletion of ATG7 or co-treatment of cells with BO-1012 and either 3-methyladenine or bafilomycin A1, two inhibitors of autophagy, not only reduced CHK1 phosphorylation and disrupted S phase arrest, but also increased cleavage of caspase-9 and PARP, and cell death. These results suggest that cells initiate S phase arrest and autophagy as pro-survival responses to BO-1012-induced DNA damage, and that suppression of autophagy enhances BO-1012-induced apoptosis via disruption of CHK1-dependent S phase arrest.

Asunto(s)

Antineoplásicos Alquilantes/farmacología; Apoptosis/efectos de los fármacos; Autofagia/efectos de los fármacos; Carcinoma/tratamiento farmacológico; Sinergismo Farmacológico; Inhibidores Enzimáticos/farmacología; Proteínas Quinasas/metabolismo; Antineoplásicos Alquilantes/agonistas; Proteína 7 Relacionada con la Autofagia; Carbamatos/agonistas; Carbamatos/farmacología; Carcinoma/enzimología; Carcinoma/metabolismo; Proliferación Celular/efectos de los fármacos; Supervivencia Celular/efectos de los fármacos; Quinasa 1 Reguladora del Ciclo Celular (Checkpoint 1); Fosfatidilinositol 3-Quinasas Clase III/antagonistas & inhibidores; Fosfatidilinositol 3-Quinasas Clase III/metabolismo; Reactivos de Enlaces Cruzados/farmacología; Femenino; Silenciador del Gen; Células HeLa; Compuestos Heterocíclicos con 3 Anillos/agonistas; Compuestos Heterocíclicos con 3 Anillos/farmacología; Humanos; Indenos/agonistas; Indenos/farmacología; Proteínas de Neoplasias/antagonistas & inhibidores; Proteínas de Neoplasias/genética; Proteínas de Neoplasias/metabolismo; Fosforilación/efectos de los fármacos; Proteínas Quinasas/química; Proteínas Quinasas/genética; Procesamiento Proteico-Postraduccional/efectos de los fármacos; Fase S/efectos de los fármacos; Enzimas Activadoras de Ubiquitina/antagonistas & inhibidores; Enzimas Activadoras de Ubiquitina/genética; Enzimas Activadoras de Ubiquitina/metabolismo; ATPasas de Translocación de Protón Vacuolares/antagonistas & inhibidores; ATPasas de Translocación de Protón Vacuolares/metabolismo

Palabras clave

Apoptosis; Autophagy; CHK1; DNA alkylating agent; DNA damage; S phase arrest

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Quinasas / Autofagia / Carcinoma / Apoptosis / Antineoplásicos Alquilantes / Sinergismo Farmacológico / Inhibidores Enzimáticos Idioma: En Revista: Toxicol Appl Pharmacol Año: 2014 Tipo del documento: Article Pais de publicación: Estados Unidos

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