Efeito do extrato de Calyptranthes grandifolia O. Berg (Myrtaceae) na expressão do TNF-α, NF-κΒ e p38α em células de Adenocarcinoma colorretal (Caco-2)
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2016-08
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Pozzobon, Adriane
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Terapias antitumorais baseadas em efeitos anti-inflamatórios vêm sendo consideradas no tratamento do câncer. A sobrevivência, proliferação e, eventualmente, invasão e metástase das células tumorais são reguladas por mediadores inflamatórios locais. Citocinas inflamatórias primárias, como o Fator de Necrose Tumoral (TNF), podem ser alvos para a terapia anticâncer. Diversos anti-inflamatórios isolados de produtos naturais vêm ganhando importância como quimiopreventivos e agentes terapêuticos para o câncer. O objetivo deste trabalho foi investigar a expressão dos genes TNF-α, NF-κΒ e p38α, associados à proliferação e inflamação em células de linhagem Caco-2 tratadas com extratos etanólico e hexânico obtidos de Calyptranthes grandifolia O.Berg (Myrtaceae). Células Caco-2 foram cultivadas e tratadas com o extrato vegetal em diferentes concentrações (200, 100, 50 e 25 μg/mL), e estimuladas com LPS. Para análise da expressão gênica foi realizada a extração de RNA total pelo método de Trizol®, seguido da síntese de DNAc (DNA complementar) e análise por qPCR (Reação em Cadeia da Polimerase em Tempo Real). Dentre os genes avaliados, observou-se uma diminuição da expressão de TNF-α nas concentrações de 100 e 200 μg/mL apenas no extrato etanólico (p<0,025, ANOVA e Teste de Tukey). O gene p38α apresentou uma tendência a aumento da expressão apenas no tratamento com extrato etanólico e o gene NF-κΒ não apresentou variações significativas em ambos os extratos analisados. Pode-se concluir que o extrato etanólico pode apresentar atividade anti-inflamatória nas concentrações testadas através da diminuição de TNF-α. Contudo, estudos complementares como análises de genotoxicidade, proteína, dentre outras, devem ser realizadas a fim de confirmar o seu potencial efeito anti-inflamatório.
Anti-tumor therapies based on anti-inflammatory effects have been considered in cancer treatment. Survival, proliferation and, eventually, invasion and metastasis of tumor cells are regulated by local inflammatory mediators. Primary inflammatory cytokines, such as Tumor Necrosis Factor (TNF), can be targets for anticancer therapy. Several anti-inflammatory isolated from natural products are acquiring importance as chemopreventive and therapeutic agents for cancer. This study aimed to investigate the expression of TNF-α, NF-κΒ and p38α genes, associated with proliferation and inflammation in Caco-2 cell line treated with ethanolic and hexanic extracts of Calyptranthes grandifolia O.Berg (Myrtaceae). Caco-2 cells were cultured and treated with plant extract at different concentrations (200, 100, 50 and 25 ug/ml) and stimulated with LPS. For gene expression analysis was performed a total RNA extraction by Trizol® method, followed by the synthesis of cDNA (complementary DNA) and analysis by qPCR (Polymerase Chain Reaction in Real Time). Among the evaluated genes, there was a decrease in TNF-α expression in 100 and 200 ug/ml concentrations only on ethanolic extract (p <0.025, ANOVA and Tukey test). The p38α gene showed a tendency to increase expression only in treatment with ethanolic extract and NF-κΒ gene did not present significant variations in both analyzed extracts. It can be concluded that ethanolic extract may present an anti-inflammatory activity on tested concentrations by decreasing TNF-α. However, further studies as genotoxicity analysis, protein, among others, should be performed to confirm its potential anti-inflammatory effect.
Anti-tumor therapies based on anti-inflammatory effects have been considered in cancer treatment. Survival, proliferation and, eventually, invasion and metastasis of tumor cells are regulated by local inflammatory mediators. Primary inflammatory cytokines, such as Tumor Necrosis Factor (TNF), can be targets for anticancer therapy. Several anti-inflammatory isolated from natural products are acquiring importance as chemopreventive and therapeutic agents for cancer. This study aimed to investigate the expression of TNF-α, NF-κΒ and p38α genes, associated with proliferation and inflammation in Caco-2 cell line treated with ethanolic and hexanic extracts of Calyptranthes grandifolia O.Berg (Myrtaceae). Caco-2 cells were cultured and treated with plant extract at different concentrations (200, 100, 50 and 25 ug/ml) and stimulated with LPS. For gene expression analysis was performed a total RNA extraction by Trizol® method, followed by the synthesis of cDNA (complementary DNA) and analysis by qPCR (Polymerase Chain Reaction in Real Time). Among the evaluated genes, there was a decrease in TNF-α expression in 100 and 200 ug/ml concentrations only on ethanolic extract (p <0.025, ANOVA and Tukey test). The p38α gene showed a tendency to increase expression only in treatment with ethanolic extract and NF-κΒ gene did not present significant variations in both analyzed extracts. It can be concluded that ethanolic extract may present an anti-inflammatory activity on tested concentrations by decreasing TNF-α. However, further studies as genotoxicity analysis, protein, among others, should be performed to confirm its potential anti-inflammatory effect.
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Inflamação; Câncer; Expressão gênica; qPCR; Calyptranthes grandifolia
Citação
DEXHEIME, Geórgia Muccillo. Efeito do extrato de Calyptranthes grandifolia O. Berg (Myrtaceae) na expressão do TNF-α, NF-κΒ e p38α em células de Adenocarcinoma colorretal (Caco-2). 2015. Dissertação (Mestrado) – Curso de Biotecnologia, Universidade do Vale do Taquari - Univates, Lajeado, 15 fev. 2015. Disponível em: http://hdl.handle.net/10737/1078.