Avaliação da resposta in vitro ao tratamento com inibidor de receptor tirosina quinase (Palladia ®) e mTORC1 (rapamicina) em células de carcinoma prostático canino
Carregando...
Data
2020-01-30
Autores
Kobayashi, Priscila Emiko [UNESP]
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
O carcinoma prostático canino possui prognóstico desfavorável e modalidades terapêuticas limitadas, principalmente em casos avançados. Devido à falta de tratamento eficaz para os carcinomas prostáticos caninos, esse estudo objetivou avaliar o efeito do toceranib (inibidor de receptor tirosina quinase) e da rapamicina (inibidor de mTORC1) em células de carcinoma prostático canino. As duas células de cultura primária advindas de carcinoma prostático canino, denominadas PC1 e PC2, apresentaram expressão proteica de VEGFR2 e PDGFR-β e ausência da proteína c-KIT, os quais são alvos do fosfato de toceranib. Adicionalmente, após comparação do transcriptoma das células não tratadas e tratadas com fosfato de toceranib, observaram-se alterações em genes relacionados com a via PDGFR, angiogênese e resistência terapêutica. Além disso, realizou-se o tratamento das células PC1 e PC2 com rapamicina (inibidor de mTORC1) após a comparação do transcriptoma das células tumorais com células de próstata normal e identificação de genes envolvidos na via PI3K/AKT/mTOR. O tratamento com rapamicina diminuiu a expressão gênica de mTOR e 4E-BP1 e aumentou de AKT, podendo representar mecanismos de resistência. Após tratamento com fosfato de toceranib, apenas a PC1 reduziu a viabilidade celular, porém após tratamento com rapamicina, as células PC1 e PC2 reduziram a viabilidade celular de maneira dose dependente, havendo resposta biológica em ambas as terapias. Sendo assim, o bloqueio de múltiplos receptores tirosina quinase e inibição de mTOR representam alvos terapêuticos no carcinoma prostático canino, entretanto a resistência terapêutica ainda é desafiadora e, portanto, a medicina de precisão seria uma forma de superar o problema.
Canine prostatic carcinoma has a poor prognosis and limited therapeutic modalities, mainly in advanced cases. The aim of this study was to evaluate the effect of toceranib phosphate (receptor tyrosine kinase inhibitor) and rapamycin (mTORC1 inhibitor) in canine prostatic carcinoma cells due to the lack of efficient therapies. Two primary canine prostate cancer cell lines, PC1 and PC2, had protein expression of VEGFR2 and PDGFR-β and absent c-KIT expression, which are target toceranib phosphate targets. Additionally, we compared transcriptome before and after toceranib phosphate treatment and we observed gene alterations related to PDGFR pathway, angiogenesis and therapeutic resistance. Besides, we treated PC1 and PC2 cells with rapamycin after transcriptome analysis comparing canine prostatic cells from normal prostate and neoplastic cells and identification of genes involved in PI3K/AKT/mTOR pathway. Gene expression of mTOR and 4E-BP1 decreased and AKT increased after rapamycin treatment, and could represent a resistency mechanism. A decrease only in PC1 cell viability was observed after toceranib phosphate treatment, however decreased cell viability in a dose-dependent manner was observed in PC1 and PC2 cells after rapamycin treatment. Therefore, we could observe biological response of canine prostatic carcinoma cells to both therapies. Thus, multiple receptor tyrosine kinase inhibition and mTOR inhibition represent target therapies to canine prostatic carcinoma. However, therapeutic resistance still remains a challenge and precision medicine could be a way to overcome this problem.
Canine prostatic carcinoma has a poor prognosis and limited therapeutic modalities, mainly in advanced cases. The aim of this study was to evaluate the effect of toceranib phosphate (receptor tyrosine kinase inhibitor) and rapamycin (mTORC1 inhibitor) in canine prostatic carcinoma cells due to the lack of efficient therapies. Two primary canine prostate cancer cell lines, PC1 and PC2, had protein expression of VEGFR2 and PDGFR-β and absent c-KIT expression, which are target toceranib phosphate targets. Additionally, we compared transcriptome before and after toceranib phosphate treatment and we observed gene alterations related to PDGFR pathway, angiogenesis and therapeutic resistance. Besides, we treated PC1 and PC2 cells with rapamycin after transcriptome analysis comparing canine prostatic cells from normal prostate and neoplastic cells and identification of genes involved in PI3K/AKT/mTOR pathway. Gene expression of mTOR and 4E-BP1 decreased and AKT increased after rapamycin treatment, and could represent a resistency mechanism. A decrease only in PC1 cell viability was observed after toceranib phosphate treatment, however decreased cell viability in a dose-dependent manner was observed in PC1 and PC2 cells after rapamycin treatment. Therefore, we could observe biological response of canine prostatic carcinoma cells to both therapies. Thus, multiple receptor tyrosine kinase inhibition and mTOR inhibition represent target therapies to canine prostatic carcinoma. However, therapeutic resistance still remains a challenge and precision medicine could be a way to overcome this problem.
Descrição
Palavras-chave
VEGFR, PDGFR, c-KIT, mTOR, Transcriptoma, Transcriptome