SET overexpression decreases cell detoxification efficiency: ALDH2 and GSTP1 are downregulated, DDR is impaired and DNA damage accumulates

Autor(es)

Almeida, Luciana O. ^[1] ; Goto, Renata N. ^[1] ; Pestana, Cezar R. ^[2] ; Uyemura, Sergio A. ^[1] ; Gutkind, Silvio ^[3] ; Curti, Carlos ^[2] ; Leopoldino, Andreia M. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Dept Anal Clin Toxicol & Bromatol, Fac Ciencias Farmaceut Ribeirao Preto, BR-14040903 Ribeirao Preto, SP - Brazil

^[2] Univ Sao Paulo, Dept Quim & Fis, Fac Ciencias Farmaceut Ribeirao Preto, BR-14040903 Ribeirao Preto, SP - Brazil

^[3] Natl Inst Dent & Craniofacial Res, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD - USA

• Tipo de documento: Artigo Científico
• Fonte: FEBS Journal; v. 279, n. 24, p. 4615-4628, DEC 2012.
• Citações Web of Science: 12

Resumo

Alcohol and tobacco consumption are risk factors for head and neck squamous cell carcinoma (HNSCC). Aldehyde dehydrogenase 2 (ALDH2) and glutathione Stransferase pi 1 (GSTP1) are important enzymes for cellular detoxification and low efficiencies are implicated in cancer. We assessed the potential role of SET protein overexpression, a histone acetylation modulator accumulated in HNSCC, in gene regulation and protein activity of ALDH2 and GSTP1. SET was knocked down in HN13, HN12 and Cal27, and overexpressed in HEK293 cells; ethanol and cisplatin were the chemical agents. Cells with SET overexpression (HEK293/SET, HN13 and HN12) showed lower ALDH2 and GSTP1 mRNA levels and trichostatin A increased them (real-time PCR). Ethanol upregulated GSTP1 and ALDH2 mRNAs, whereas cisplatin upregulated GSTP1 in HEK293 cells. SET-chromatin binding revealed SET interaction with ALDH2 and GSTP1 promoters, specifically via SET NAP domain; ethanol and cisplatin abolished SET binding. ALDH2 and GSTP1 efficiency was assessed by enzymatic and comet assay. A lower ALDH2 activity was associated with greater DNA damage (tail intensity) in HEK293/SET compared with HEK293 cells, whereas HN13/siSET showed ALDH2 activity higher than HN13 cells. HN13/siSET cells showed increased tail intensity. Cisplatin-induced DNA damage response showed negative relationship between SET overexpression and BRCA2 recruitment. SET downregulated repair genes ATM, BRCA1 and CHEK2 and upregulated TP53. Cisplatin-induced cell-cycle arrest occurred in G0/G1 and S in HEK293 cells, whereas HEK293/SET showed G2/M stalling. Overall, cisplatin was more cytotoxic for HN13 than HN13/siSET cells. Our data suggest a role for SET in cellular detoxification, DNA damage response and genome integrity. (AU)

• Processo FAPESP: 10/08328-7 - Estudos da resistência à apoptose induzida por t-butil hidroperoxido no câncer, modelo cabeça/pescoço: vias de sinalização com ênfase na PIP3-Akt/SET, estresse oxidativo, mitocôndria e correlações
• Beneficiário: Cezar Rangel Pestana
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 10/20536-4 - Estudo das funções da região ácida e domínio NAP da proteína SET em linhagem celular
• Beneficiário: Renata Nishida Goto
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 09/52228-0 - Estudos da resistência à apoptose no câncer, modelo cabeça/pescoço: vias de sinalização com ênfase na PIP3-AKT/SET, estresse oxidativo, mitocôndria e correlações
• Beneficiário: Carlos Curti
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 09/10783-7 - Estudos moleculares do envolvimento das proteínas hnRNP K e SET na transcrição e tradução de proteínas associadas à tumorigênese oral
• Beneficiário: Luciana Oliveira de Almeida
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 10/20384-0 - Identificação dos genes, miRNAs e proteínas regulados pela oncoproteína SET e envolvidos na malignização e progressão do carcinoma epidermóide de cabeça e pescoço usando modelo in vitro e in vivo
• Beneficiário: Andréia Machado Leopoldino
• Modalidade de apoio: Auxílio à Pesquisa - Regular