Atividade do micronutriente selênio na cinética de proliferação celular, citotoxicidade, indução de apoptose e expressão dos genes CASP9, BCL-XL e APC em células HT29
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Data
2011-02-17
Autores
Mauro, Mariana de Oliveira [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Cânceres intestinais são correlacionados com a dieta. Assim, a determinação e compreensão das interações nutriente-genoma é importante. O presente trabalho avaliou a ação do selênio oligoelemento na citotoxicidade de proliferação celular, e na indução de apoptose in situ e na expressão dos genes CASP9, BCL-XL e APC em células de adenocarcinoma intestinal (HT29). Células HT29 foram cultivadas e tratadas com selênio nas concentrações de 5, 50 e 500 ng/mL, com ou sem o agente indutor de danos doxorrubicina. Essas células foram então avaliadas por meio do ensaio de citotoxicidade (MTT), proliferação celular e na indução de apoptose in situ. Para avaliar a expressão gênica, apenas as células tratadas com 500 ng/mL de selênio foram utilizadas. O RNA foi extraído a partir dessas células, e as expressões de CASP9, BCL-XL e APC foram analisadas pelo método de RTPCR. O gene GAPDH foi utilizado como gene de referência. O MTT mostrou que o selênio não foi tóxico em qualquer uma das concentrações testadas. O ensaio de proliferação celular mostrou que o selênio não interfere com a proliferação celular nas três concentrações testadas. Em contraste, quando as três concentrações foram combinadas com a doxorrubicina, uma diminuição significativa na taxa de proliferação celular foi observada. A taxa de apoptose foi significativamente maior no selênio (500 ng/mL) e doxorrubicina. A expressão CASP9 foi aumentada e expressão BCL-XL diminuiu no selênio (500 ng/mL) e o grupo de doxorrubicina. A expressão do gene APC foi significativamente maior no grupo selênio sozinho. Estes resultados mostram que a apoptose selênio aumenta, especialmente quando está associado com um agente de indução de danos. Além disso, o selênio tem um papel importante na expressão do gene APC, que está relacionada com a regulação do ciclo celular
Intestinal cancers are correlated with diet. Thus, determining and understanding nutrient-genome interactions is important. The present work assessed the action of the oligoelement selenium on cell proliferation, cytotoxicity, and in situ apoptosis induction and on the expression CASP9, BCL-XL and APC genes in intestinal adenocarcinoma cells (HT29). HT29 cells were cultured and treated with selenium at concentrations of 5, 50 and 500 ng/mL with or without the damage-inducing agent doxorubicin. These cells were then evaluated for cytotoxicity (MTT), cell proliferation and in situ apoptosis induction. To evaluate gene expression, only the cells treated with 500 ng/mL of selenium were used. RNA was extracted from these cells, and the expressions of CASP9, BCL-XL and APC were analyzed by the RT-PCR method. The GAPDH gene was used as a reference gene. The MTT assay showed that selenium was not cytotoxic at any of the concentrations tested. The cell proliferation assay showed that selenium did not interfere with cell proliferation at the three concentrations tested. In contrast, when the three concentrations were combined with doxorubicin, a significant decrease in the proliferation rate was observed. The apoptosis rate was significantly increased in the selenium (500 ng/mL) and doxorubicin group. CASP9 expression was increased and BCL-XL expression decreased in the selenium (500 ng/mL) and doxorubicin group. APC was significantly increased in the selenium group alone. These results show that selenium increases apoptosis, especially when it is associated with a damage-inducing agent. Also, selenium has an important role in the expression of the APC gene, which is related to cell cycle regulation
Intestinal cancers are correlated with diet. Thus, determining and understanding nutrient-genome interactions is important. The present work assessed the action of the oligoelement selenium on cell proliferation, cytotoxicity, and in situ apoptosis induction and on the expression CASP9, BCL-XL and APC genes in intestinal adenocarcinoma cells (HT29). HT29 cells were cultured and treated with selenium at concentrations of 5, 50 and 500 ng/mL with or without the damage-inducing agent doxorubicin. These cells were then evaluated for cytotoxicity (MTT), cell proliferation and in situ apoptosis induction. To evaluate gene expression, only the cells treated with 500 ng/mL of selenium were used. RNA was extracted from these cells, and the expressions of CASP9, BCL-XL and APC were analyzed by the RT-PCR method. The GAPDH gene was used as a reference gene. The MTT assay showed that selenium was not cytotoxic at any of the concentrations tested. The cell proliferation assay showed that selenium did not interfere with cell proliferation at the three concentrations tested. In contrast, when the three concentrations were combined with doxorubicin, a significant decrease in the proliferation rate was observed. The apoptosis rate was significantly increased in the selenium (500 ng/mL) and doxorubicin group. CASP9 expression was increased and BCL-XL expression decreased in the selenium (500 ng/mL) and doxorubicin group. APC was significantly increased in the selenium group alone. These results show that selenium increases apoptosis, especially when it is associated with a damage-inducing agent. Also, selenium has an important role in the expression of the APC gene, which is related to cell cycle regulation
Descrição
Palavras-chave
Cancer, Selenio, Celulas - Proliferação, Intestino grosso - Câncer, Citotoxicidade, Apoptose, Antioxidantes, Carcinogenos, Anticarcinogenicos, Cancer - Aspectos nutricionais
Como citar
MAURO, Mariana de Oliveira. Atividade do micronutriente selênio na cinética de proliferação celular, citotoxicidade, indução de apoptose e expressão dos genes CASP9, BCL-XL e APC em células HT29. 2011. 82 f. Dissertação - (mestrado) - Universidade Estadual Paulista, Instituto de Biociências de Rio Claro, 2011.