Avaliação da terapia associada de peptídeos derivados do inibidor de proteases EcTI com paclitaxel e doxorrubicina em linhagem de câncer de mama triplo-negativo
Data
2023-01-12
Autores
Rocha, Vitoria Morais da [UNIFESP]
Orientadores
Oliva, Maria Luiza Vilela [UNIFESP]
Tipo
Trabalho de conclusão de curso de graduação
Título da Revista
ISSN da Revista
Título de Volume
Resumo
O câncer de mama triplo negativo é um subtipo que não apresenta receptores para estrogênio, progesterona, e nem expressa HER-2, dificultando a terapia alvo e a terapia hormonal. Os dois quimioterápicos mais utilizados no tratamento deste subtipo são o paclitaxel (PTX) e a doxorrubicina (DOX), entretanto, é conhecida a significante ocorrência de efeitos colaterais e quimiorresistência, reforçando a necessidade de tratamentos alternativos e/ou adjuvantes. No câncer há um aumento da expressão de proteases, que contribuem com o sucesso da progressão tumoral. Assim sendo, inibidores de protease têm se tornado uma alternativa terapêutica, e o EcTI (Inibidor de Tripsina de Enterolobium contortisiliquum), apresentou efeito antitumoral em estudos preliminares, inclusive no câncer de mama triplo-negativo. Peptídeos são fortes candidatos à terapia, combinados à terapia com dois ou mais fármacos. Por esta razão, avaliamos o efeito de peptídeos sintéticos (PEP 2, PEP 3 e PEP 7) derivados da sequência do EcTI, bem como sua associação ao PTX e a DOX, como possível alternativa de redução de efeitos colaterais e tratamento para o câncer de mama triplo-negativo. A avaliação do efeito antitumoral foi realizada por meio de ensaios celulares de viabilidade, migração, morte celular, ciclo celular e proliferação, além da análise da degradação dos peptídeos e western blot. Os resultados avaliados por ANOVA de uma via mostraram que o PEP 2 e PEP 7 reduziram a viabilidade nos tempos investigados, com destaque para o PEP 7. Os peptídeos não afetaram as células da glândula mamária, indicando sua seletividade para as células tumorais. Além disso, os peptídeos inibiram a migração celular via bloqueio de metalo-proteases, induziram a morte celular por apoptose e inibiram a proliferação celular. Não foi observada a potencialização do efeito com a terapia associada, no entanto, os peptídeos isoladamente foram capazes de induzir efeito antitumoral eficazmente. Os resultados obtidos são relevantes, contribuem com a busca por novos compostos para o tratamento do subtipo triplo-negativo e impulsionam para a continuidade dos estudos para a identificação dos mecanismos responsáveis pelos efeitos dos peptídeos.
Triple-negative breast cancer is a subtype that does not have receptors for estrogen, and progesterone, and does not express HER-2, making targeted therapy and hormone therapy difficult. The two most used chemotherapeutics in the treatment of this subtype are paclitaxel (PTX) and doxorubicin (DOX), however, the significant occurrence of side effects and chemoresistance is known, reinforcing the need for alternative and/or adjuvant treatments. In cancer, there is an increase in the expression of proteases, which contribute to the success of tumor progression. Therefore, protease inhibitors are a therapeutic alternative, and EcTI (Enterolobium contortisiliquum Trypsin Inhibitor), showed an antitumor effect in preliminary studies, including in triple-negative breast cancer. Peptides are strong candidates for combined therapy with two or more drugs. For this reason, we evaluated the effect of synthetic peptides (PEP 2, PEP 3, and PEP 7) derived from the EcTI sequence, as well as their association with PTX and DOX, as a possible alternative for reducing side effects and treating triple-negative breast cancer. The evaluation of the antitumor effect was carried out through cell viability, migration, cell death, cell cycle, and proliferation assays, in addition to the analysis of peptide degradation and western blot. The results evaluated by One-way ANOVA showed that PEP 2 and PEP 7 reduced cell viability at the investigated times, with emphasis on PEP 7. The peptides did not affect mammary gland cells, indicating their selectivity for tumor cells. Furthermore, the peptides inhibited cell migration by blocking metalloproteases, induced cell death by apoptosis and inhibited cell proliferation. No potentiation of the effect was observed with the associated therapy; however, the peptides alone were able to induce an antitumor effect. The results obtained are relevant, contribute to the search for new compounds for the treatment of the triple-negative subtype, and encourage the continuation of studies to identify the mechanisms responsible for the effects of the peptides.
Triple-negative breast cancer is a subtype that does not have receptors for estrogen, and progesterone, and does not express HER-2, making targeted therapy and hormone therapy difficult. The two most used chemotherapeutics in the treatment of this subtype are paclitaxel (PTX) and doxorubicin (DOX), however, the significant occurrence of side effects and chemoresistance is known, reinforcing the need for alternative and/or adjuvant treatments. In cancer, there is an increase in the expression of proteases, which contribute to the success of tumor progression. Therefore, protease inhibitors are a therapeutic alternative, and EcTI (Enterolobium contortisiliquum Trypsin Inhibitor), showed an antitumor effect in preliminary studies, including in triple-negative breast cancer. Peptides are strong candidates for combined therapy with two or more drugs. For this reason, we evaluated the effect of synthetic peptides (PEP 2, PEP 3, and PEP 7) derived from the EcTI sequence, as well as their association with PTX and DOX, as a possible alternative for reducing side effects and treating triple-negative breast cancer. The evaluation of the antitumor effect was carried out through cell viability, migration, cell death, cell cycle, and proliferation assays, in addition to the analysis of peptide degradation and western blot. The results evaluated by One-way ANOVA showed that PEP 2 and PEP 7 reduced cell viability at the investigated times, with emphasis on PEP 7. The peptides did not affect mammary gland cells, indicating their selectivity for tumor cells. Furthermore, the peptides inhibited cell migration by blocking metalloproteases, induced cell death by apoptosis and inhibited cell proliferation. No potentiation of the effect was observed with the associated therapy; however, the peptides alone were able to induce an antitumor effect. The results obtained are relevant, contribute to the search for new compounds for the treatment of the triple-negative subtype, and encourage the continuation of studies to identify the mechanisms responsible for the effects of the peptides.