Estudo da ação da emodina associada à terapia fotodinâmica em células de carcinoma cervical positivas para HPV de alto risco
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Data
2019-12-16
Autores
Campoy, Ana Emília Brumatti Galiardi
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O câncer cervical é um problema de saúde pública mundial, especialmente devido à sua relação com a infecção prévia pelo papilomavírus humano (HPV). Os HPV são uma família de vírus de DNA com mais de 200 tipos e podem ser classificados em HPVs de baixo e alto risco. Os HPVs de alto risco mais relevantes são os HPV-16 e -18, que juntos são responsáveis por mais de 70% dos casos de carcinoma cervical. As modalidades atuais de tratamento para o câncer cervical são cirurgia, e a combinação de quimioterapia à base de cisplatina com radiação, porém apresentam efeitos adversos graves. Portanto, esforços contínuos são necessários para desenvolver novas drogas e estratégias terapêuticas eficazes para aumentar a eficácia da quimioterapia e diminuir esses efeitos colaterais. A emodina tem atraído grande atenção devido a seu efeito anti-inflamatório, antineoplásico e proapoptótico nos últimos anos. Além disso, a emodina pode ser utilizada como agente fotossensibilizador na terapia fotodinâmica. O interesse na terapia fotodinâmica no tratamento do câncer tem crescido exponencialmente, uma vez que é um tratamento minimamente invasivo, onde se erradica as células alvo, evitando-se a toxicidade sistêmica e os efeitos colaterais nos tecidos saudáveis. Assim, o objetivo deste estudo foi analisar o efeito da emodina associada à terapia fotodinâmica em linhagens de carcinoma cervical infectadas por HPV de alto risco (SiHa e CaSki) e queratinócitos humanos imortalizados (HaCaT). Inicialmente, as investigações demonstraram que a emodina apresentou citotoxicidade em concentração e tempo dependentes, mostrando mais de 80% de viabilidade celular para linhagens celulares SiHa, CaSki e HaCaT em concentrações abaixo de 30 µmol/L. As imagens de microscopia de fluorescência mostraram uma internalização celular eficiente de emodina em todas as linhas celulares analisadas. Uma diminuição significativa da viabilidade celular para as linhagens celulares SiHa, CaSki e HaCaT foi observado após tratamento com emodina associada à terapia fotodinâmica. Isto foi acompanhado por um aumento na produção de EROs, aumento nas atividades de caspase-3 e aumento na intensidade de fluorescência dos vacúolos autofágicos, sugerindo morte celular por apoptose e autofagia, provavelmente devido ao aumento na produção de EROs. Além disso, 22 genes alvo de drogas anticâncer foram superexpressos e 2 genes alvo de drogas anticâncer tiveram sua expressão diminuída em células SiHa após o tratamento com terapia fotodinâmica mediada por emodina. Estes resultados mostram que a emodina em combinação com a terapia fotodinâmica tem um grande potencial terapêutico para ser utilizada como um tratamento alternativo para o câncer cervical.
Cervical cancer is a worldwide public health problem, especially due to its relation to the previous infection by human papillomavirus (HPV). The HPV is a family of DNA viruses with more than 200 types, and can be classified in low and high risk HPVs. The most important high risk HPVs are the HPV-16 and -18, which together are responsible for more than 70% cervical carcinoma cases. Current treatment modalities for cervical cancer are surgery, and the combination of cisplatin based chemotherapy with radiation, however they present severe adverse effects. Therefore, ongoing efforts are necessary to developed new drugs and effective therapeutic strategies to enhance chemotherapeutic efficacy and decrease these side effects. Emodin has attracted extensive attention due to its anti-inflammatory, antineoplastic, and proapoptotic effects in recent years. Furthermore, emodin may be used as a photosensitizing agent in photodynamic therapy. The interest in photodynamic therapy in the treatment of cancer has grown exponentially, since it is a minimally invasive treatment where eradicate target cells while avoiding systemic toxicity and side effects on healthy tissues. So, the aim of this study was to analyze the effect of emodin associated with photodynamic therapy in cervical carcinoma cell lines infected with high-risk HPV (SiHa and CaSki) and immortalized human keratinocytes (HaCaT). Initially, the investigations demonstrated that emodin presented cytotoxicity in concentration and time-dependent manner showing more than 80% of cell viability for SiHa, CaSki and HaCaT cell lines in concentrations below 30 µmol/L. The fluorescence microscopy images showed an efficient cellular uptake of emodin in all the cell lines analyzed. A significant decrease of the cell viability for SiHa, CaSki and HaCaT cell lines was observed after treatment with photodynamic therapy mediated by emodin. This was accompanied by an increase of ROS production, increase in caspase-3 activity and increase in the fluorescence intensity of autophagic vacuoles, suggesting cell death by apoptosis and autophagy probably due to the increase in production of ROS. Additionally, 22 target genes for anti-cancer drugs were overexpressed and 2 target genes for anti-cancer drugs were downregulated in SiHa cells after treatment with photodynamic therapy mediated by emodin. These results show that emodin in combination with photodynamic therapy has a great therapeutic potential to be used as an alternative treatment for cervical cancer.
Cervical cancer is a worldwide public health problem, especially due to its relation to the previous infection by human papillomavirus (HPV). The HPV is a family of DNA viruses with more than 200 types, and can be classified in low and high risk HPVs. The most important high risk HPVs are the HPV-16 and -18, which together are responsible for more than 70% cervical carcinoma cases. Current treatment modalities for cervical cancer are surgery, and the combination of cisplatin based chemotherapy with radiation, however they present severe adverse effects. Therefore, ongoing efforts are necessary to developed new drugs and effective therapeutic strategies to enhance chemotherapeutic efficacy and decrease these side effects. Emodin has attracted extensive attention due to its anti-inflammatory, antineoplastic, and proapoptotic effects in recent years. Furthermore, emodin may be used as a photosensitizing agent in photodynamic therapy. The interest in photodynamic therapy in the treatment of cancer has grown exponentially, since it is a minimally invasive treatment where eradicate target cells while avoiding systemic toxicity and side effects on healthy tissues. So, the aim of this study was to analyze the effect of emodin associated with photodynamic therapy in cervical carcinoma cell lines infected with high-risk HPV (SiHa and CaSki) and immortalized human keratinocytes (HaCaT). Initially, the investigations demonstrated that emodin presented cytotoxicity in concentration and time-dependent manner showing more than 80% of cell viability for SiHa, CaSki and HaCaT cell lines in concentrations below 30 µmol/L. The fluorescence microscopy images showed an efficient cellular uptake of emodin in all the cell lines analyzed. A significant decrease of the cell viability for SiHa, CaSki and HaCaT cell lines was observed after treatment with photodynamic therapy mediated by emodin. This was accompanied by an increase of ROS production, increase in caspase-3 activity and increase in the fluorescence intensity of autophagic vacuoles, suggesting cell death by apoptosis and autophagy probably due to the increase in production of ROS. Additionally, 22 target genes for anti-cancer drugs were overexpressed and 2 target genes for anti-cancer drugs were downregulated in SiHa cells after treatment with photodynamic therapy mediated by emodin. These results show that emodin in combination with photodynamic therapy has a great therapeutic potential to be used as an alternative treatment for cervical cancer.
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Palavras-chave
Câncer cervical, Emodina, Papilomavírus humano, Terapia fotodinâmica, Cervical cancer, Emodin, Human papillomavirus, Photodynamic therapy