Efeitos da melatonina nas células de carcinoma ovariano SKOV-3: análise do perfil proteômico global
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Data
2022-01-26
Autores
Cesário, Roberta Carvalho
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Universidade Estadual Paulista (Unesp)
Resumo
O câncer de ovário (CO) é a segunda malignidade mais letal do sistema genital feminino e apresenta uma expectativa de sobrevida menor que 50% após o quinto ano do diagnóstico. Dependendo do subtipo, o CO está associado com maior risco de desenvolvimento de quimioresistência e prognóstico ruim. A melatonina (Mel) é um neurohormônio secretado pela glândula pineal na fase de escuro e tem mostrado propriedades antitumorais em ensaios in vitro e in vivo. O objetivo do presente estudo foi avaliar a influência da terapia com melatonina, em diferentes doses, sobre o proteoma das células de carcinoma ovariano humano (linhagem SKOV-3). O estudo foi composto pelos seguintes grupos experimentais; 1) Grupo Controle: células SKOV-3 tratadas com veículo; 2) Grupo Mel 0.8 mM: células SKOV-3 tratadas com melatonina na concentração de 0.8 mM por 48h; 3) Grupo Mel 1.6 mM: células SKOV-3 tratadas com melatonina na concentração de 1.6 mM por 48h; 4) Grupo Mel 2.4 mM: células SKOV-3 tratadas com melatonina na concentração de 2.4 mM por 48h. Após 48h de tratamento com melatonina, foi realizada a extração e a quantificação das proteínas. O perfil proteômico foi analisado e identificou-se 636 proteínas totais no grupo controle. A partir desses dados, foram selecionadas proteínas diferencialmente reguladas para cada grupo, comparando as expressões das proteínas do grupo controle usando p<0,05 e fold change de +1.5 ou -1.5 para determinar as proteínas que foram reguladas de maneira positiva e negativa, respectivamente. Proteínas relacionadas com o sistema imune e o metabolismo do ácido tricarboxílico foram reguladas positivamente nos três grupos expostos a melatonina. Especificamente, a dose de 2.4 mM promoveu alterações importantes, regulando genes envolvidos na síntese de proteínas. Também identificamos 28 genes potenciais compartilhados entre o tecido normal e o tecido tumoral ovariano considerando todos os grupos experimentais, onde a melatonina mostrou alterações na predição de genes codificadores de proteínas ribossomais. Concluímos que a melatonina modula o proteoma das células SKOV-3, principalmente regulando proteínas envolvidas com o sistema imune e metabolismo mitocondrial. Além disso, o tratamento com a dose de 2.4 mM de melatonina promoveu maiores alterações na rede proteica, revelando uma estratégia terapêutica eficaz no combate ao CO.
Ovarian cancer (OC) is the second most lethal malignancy of the female genital system and has a survival expectation of less than 50% after the fifth year diagnosis. Depending on the subtype, OC is associated with an increased risk of chemoresistance and poor prognosis. Melatonin (5- methoxytriptamine) is a neurohormone secreted by the pineal gland in the dark phase and has shown antitumor properties in both in vitro and in vivo assays. The present study aimed to evaluate the influence of melatonin therapy, at different doses, on the proteome of human ovarian carcinoma cells (SKOV-3 cell line). The study consisted of the following experimental groups; 1) Control Group: SKOV-3 cells treated with vehicle; 2) 0.8 mM Mel Group: SKOV3 cells treated with melatonin at a concentration of 0.8 mM for 48h; 3) Mel 1.6 mM Group: SKOV-3 cells treated with melatonin at a concentration of 1.6 mM for 48h; 4) Mel 2.4 mM group: SKOV-3 cells treated with melatonin at a concentration of 2.4 mM for 48h. After 48 h of treatment with melatonin, protein extraction and quantification were performed in these cells. The proteomic profile was analyzed, and a total of 636 proteins were identified in the control group. From these data, differentially regulated proteins were selected for each group, comparing the level of the proteins in the control group using p<0.05 and fold change of +1.5 or -1.5 to determine which proteins were positively and negatively regulated, respectively. Proteins related to the immune system and tricarboxylic acid metabolism were upregulated in the three groups exposed to melatonin. More specifically, the dose of 2.4 mM promoted significant changes, thereby regulating molecules involved in protein synthesis. We also identified 28 potential genes shared between normal tissue and ovarian tumor tissue considering all experimental groups, where melatonin showed alterations in predicting genes encoding ribosomal proteins. We conclude that melatonin modulates the proteome of SKOV-3 cells, mainly via regulating proteins involved with the immune system and mitochondrial metabolism. Furthermore, treatment with a dose of 2.4 mM of melatonin promoted more significant alterations in the protein network, thus revealing an effective therapeutic strategy against OC.
Ovarian cancer (OC) is the second most lethal malignancy of the female genital system and has a survival expectation of less than 50% after the fifth year diagnosis. Depending on the subtype, OC is associated with an increased risk of chemoresistance and poor prognosis. Melatonin (5- methoxytriptamine) is a neurohormone secreted by the pineal gland in the dark phase and has shown antitumor properties in both in vitro and in vivo assays. The present study aimed to evaluate the influence of melatonin therapy, at different doses, on the proteome of human ovarian carcinoma cells (SKOV-3 cell line). The study consisted of the following experimental groups; 1) Control Group: SKOV-3 cells treated with vehicle; 2) 0.8 mM Mel Group: SKOV3 cells treated with melatonin at a concentration of 0.8 mM for 48h; 3) Mel 1.6 mM Group: SKOV-3 cells treated with melatonin at a concentration of 1.6 mM for 48h; 4) Mel 2.4 mM group: SKOV-3 cells treated with melatonin at a concentration of 2.4 mM for 48h. After 48 h of treatment with melatonin, protein extraction and quantification were performed in these cells. The proteomic profile was analyzed, and a total of 636 proteins were identified in the control group. From these data, differentially regulated proteins were selected for each group, comparing the level of the proteins in the control group using p<0.05 and fold change of +1.5 or -1.5 to determine which proteins were positively and negatively regulated, respectively. Proteins related to the immune system and tricarboxylic acid metabolism were upregulated in the three groups exposed to melatonin. More specifically, the dose of 2.4 mM promoted significant changes, thereby regulating molecules involved in protein synthesis. We also identified 28 potential genes shared between normal tissue and ovarian tumor tissue considering all experimental groups, where melatonin showed alterations in predicting genes encoding ribosomal proteins. We conclude that melatonin modulates the proteome of SKOV-3 cells, mainly via regulating proteins involved with the immune system and mitochondrial metabolism. Furthermore, treatment with a dose of 2.4 mM of melatonin promoted more significant alterations in the protein network, thus revealing an effective therapeutic strategy against OC.
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Palavras-chave
Câncer de ovário, Melatonina, Proteoma, Mitocondria, Células SKOV-3, Ovarian cancer, Melatonin, Proteome, Mitochondria