Efeitos da melatonina sobre a proliferação induzida pela norepinefrina e epinefrina em células de câncer de boca.
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Data
2020-11-30
Autores
Coachman, Laura Ceretti [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O presente estudo teve por objetivo avaliar a regulação da melatonina (MT) sobre proliferação celular e expressão de interleucina-6 (IL-6) em células de câncer de boca induzida pela norepinefrina (NE) e epinefrina (EP). Primeiramente, células das linhagens SCC9 e SCC25 foram cultivadas em meio de cultura composto de Eagle modificado por Dulbecco’s (DMEM) e meio de cultura F12 (DMEM/F12) e mantidas a 37°C em atmosfera úmida de 5% de gás carbônico (CO2). Os estímulos hormonais foram realizados por 3, 6 e 24h com MT em concentrações de 1mM, 5mM e 10mM, EP 10 μM e NE 10 μ. Para os grupos em que se avaliou o efeito da MT com os hormônios do estresse, as células foram estimuladas com MT (1mM, 5mM e 10mM) 1 hora antes dos estímulos com EP (10 μM) e NE (10 μM). Células não estimuladas foram utilizadas como controle. O potencial proliferativo das células foi avaliado pela técnica de BrdU com kit específico e por ensaio de MTT. A expressão de IL-6 foi analisada por rt-PCR em tempo real. Os resultados mostraram que os hormônios epinefrina e norepinefrina induziram aumento da proliferação das células SCC9 e SCC25 em todos os períodos avaliados. A melatonina em concentrações de 5 e 10 mM promoveu redução da proliferação celular induzida pela epinefrina e norepinefrina. A expressão gênica de IL-6 mostrou-se aumentada após 6 horas de tratamento com norepinefrina e epinefrina (10 μM). O tratamento com melatonina (5 mM) foi capaz de reduzir significativamente a expressão de IL-6 induzida pelos hormônios do estresse. Estes resultados em conjunto denotam que a melatonina pode atenuar a proliferação de células de câncer de boca induzida por hormônios relacionados ao estresse, efeito este que pode ter sido associado à influência da melatonina sobre a expressão de IL-6.
The present study aimed to evaluate the regulation of melatonin on cell proliferation and IL-6 expression by norepinephrine and epinephrine-induced oral cancer cells. First, SCC9 and SCC25, cell strains were cultured in Dulbecco's modified Eagle's compound culture medium (DMEM) and F12 culture medium (DMEM / F12) and maintained at 37 ° C in a humidified 5% CO2 atmosphere. Hormonal stimuli were performed with norepinephrine (10 μM) and epinephrine (10 μM) for 3, 6 and 24h in the presence or absence of melatonin (1 mM, 5 mM and 10 mM). Unstimulated cells were used as controls. Cell proliferative potential was evaluated by BrdU technique with specific kit and MTT assay. IL-6 expression was analyzed by rt-PCR in real time. The results showed that the epinephrine and norepinephrine hormones induced significant increase of SCC9 and SCC25 cell proliferation in all periods evaluated. Melatonin at concentrations of 5 and 10 mM demonstrated potential to reduce epinephrine and norepinephrine-induced cell proliferation. IL-6 gene expression was significantly increased after treatment with norepinephrine and epinephrine (10 μM) after 6 hours. Treatment with melatonin (5 mM) in the presence of epinephrine or norepinephrine was able to significantly reduce IL-6 expression. These findings showed that melatonin may affect the proliferation of oral cancer cells induced by stress-related hormones (norepinephrine and epinephrine) and this effect was mediated by IL-6 expression.
The present study aimed to evaluate the regulation of melatonin on cell proliferation and IL-6 expression by norepinephrine and epinephrine-induced oral cancer cells. First, SCC9 and SCC25, cell strains were cultured in Dulbecco's modified Eagle's compound culture medium (DMEM) and F12 culture medium (DMEM / F12) and maintained at 37 ° C in a humidified 5% CO2 atmosphere. Hormonal stimuli were performed with norepinephrine (10 μM) and epinephrine (10 μM) for 3, 6 and 24h in the presence or absence of melatonin (1 mM, 5 mM and 10 mM). Unstimulated cells were used as controls. Cell proliferative potential was evaluated by BrdU technique with specific kit and MTT assay. IL-6 expression was analyzed by rt-PCR in real time. The results showed that the epinephrine and norepinephrine hormones induced significant increase of SCC9 and SCC25 cell proliferation in all periods evaluated. Melatonin at concentrations of 5 and 10 mM demonstrated potential to reduce epinephrine and norepinephrine-induced cell proliferation. IL-6 gene expression was significantly increased after treatment with norepinephrine and epinephrine (10 μM) after 6 hours. Treatment with melatonin (5 mM) in the presence of epinephrine or norepinephrine was able to significantly reduce IL-6 expression. These findings showed that melatonin may affect the proliferation of oral cancer cells induced by stress-related hormones (norepinephrine and epinephrine) and this effect was mediated by IL-6 expression.
Descrição
Palavras-chave
Melatonina, Neoplasias bucais, Stress (Psicologia), Norepinefrina, Adrenalina, Interleucina-6, Mouth neoplasms