Estudo comparado dos efeitos antitumorais dos extratos de cannabis sativa rico em thc e cbd em células de carcinoma prostático canino e humano
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Data
2023-06-23
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Universidade Estadual Paulista (Unesp)
Resumo
O carcinoma de próstata é o segundo tipo de neoplasia maligna mais comum entre os homens no Brasil, e relatado em cães entre 8 a 10 anos de idade, com rápida evolução podendo metastatizar para linfonodos regionais, pulmões e ossos. O CBD já foi relatado na literatura como agente antineoplásico em modelos in vitro de neoplasias do homem e do cão. Nós investigamos o efeito citotóxico dos extratos de dois extratos de Cannabis sativa, um rico em CBD (ERCBD) e outro rico em THC (ERTHC), em linhagens de células de carcinoma prostático canino (PC1 e PC2) e humano (LNCAP), bem como sua avaliação para a indução a apoptose de células cancerígenas. Para avaliação da citotoxicidade foi o utilizado o ensaio de MTT, com as doses 2,5µM, 5µM, 7,5µM e 10µM de ambos os compostos nas linhagens caninas e 10µM, 20µM, 30µM e 40µM de ambos os compostos na linhagem humana, As linhagens foram incubadas por 24 horas, em placas de 96 poços em condições de cultura in vitro padrão. Após esse período determinou-se o IC50 através da atividade metabólica das células. Logo após foi realizado o ensaio da Anexina-V-FITC/ Iodeto de propídio nas linhagens caninas, avaliando através da citometria de fluxo as células com apoptose mediata e tardia. Com os resultados obtidosobtiveram uma concentração inibitória média de ERCBD (3,437; 3,577) e ERTHC (4,909;4,486) em PC1 e PC2 respectivamente, e ERCBD (21,49) e ERTHC (18,69) na linhagem LNCAP. Ainda, foi realizado ensaio de apoptose nas linhagens de carcinoma prostático canino, após o tratamento. Mais estudos devem ser feitos, para avaliar o mecanismo de morte celular e seu potencial de uso in vivo.
Prostate carcinoma is the second most common type of cancer among men in Brazil, and is reported in dogs between 8 and 10 years of age, with rapid evolution and metastatic potential to regional lymph nodes, lungs and bones. CBD and THC showed an antitumoral effect in human and canine cell lines. We investigated the cytotoxic effect of extracts from two Cannabis sativa extracts, one rich in CBD (ERCBD) and the other rich in THC (ERTHC), on malignant prostatic cell lines from dogs (PC1 and PC2) and men (LNCAP), as well as the effect evaluation for the induction of apoptosis in the two canine cell lines. To evaluate the cytotoxicity, the MTT assay was used, with doses of 2.5µM, 5µM, 7.5µM and 10µM of both compounds in the canine cells line and 10µM, 20µM, 30µM and 40µM of both compounds in the human cell line.Both were incubated for 24 hours in 96-well plates. The IC50 was determined through the metabolic activity of the cells. Soon after, the Annexin-V-FITC/propidium iodide assay was performed in the canine PC cell lines, evaluating cells with mediate and late apoptosis through flow cytometry. We showed that ERCBD and ERTHC obtained an average inhibitory concentration of 3.437; 3.577 and 4.909;4.486 in PC1 and PC2 respectively, and ERCBD (21.49) and ERTHC (18 ,69) in the LNCAP cell line. , We also observed an apoptotic effect on the canine cell lines for both cannabis extracts. Further research should be made to evaluate the cell death mechanism as well as the in vitro use as an anti neoplastic agent.
Prostate carcinoma is the second most common type of cancer among men in Brazil, and is reported in dogs between 8 and 10 years of age, with rapid evolution and metastatic potential to regional lymph nodes, lungs and bones. CBD and THC showed an antitumoral effect in human and canine cell lines. We investigated the cytotoxic effect of extracts from two Cannabis sativa extracts, one rich in CBD (ERCBD) and the other rich in THC (ERTHC), on malignant prostatic cell lines from dogs (PC1 and PC2) and men (LNCAP), as well as the effect evaluation for the induction of apoptosis in the two canine cell lines. To evaluate the cytotoxicity, the MTT assay was used, with doses of 2.5µM, 5µM, 7.5µM and 10µM of both compounds in the canine cells line and 10µM, 20µM, 30µM and 40µM of both compounds in the human cell line.Both were incubated for 24 hours in 96-well plates. The IC50 was determined through the metabolic activity of the cells. Soon after, the Annexin-V-FITC/propidium iodide assay was performed in the canine PC cell lines, evaluating cells with mediate and late apoptosis through flow cytometry. We showed that ERCBD and ERTHC obtained an average inhibitory concentration of 3.437; 3.577 and 4.909;4.486 in PC1 and PC2 respectively, and ERCBD (21.49) and ERTHC (18 ,69) in the LNCAP cell line. , We also observed an apoptotic effect on the canine cell lines for both cannabis extracts. Further research should be made to evaluate the cell death mechanism as well as the in vitro use as an anti neoplastic agent.
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Canabinoide, Cães - Doenças, Próstata - Câncer, Canabinóides, Medicamento fitoterápico, Maconha, Cannabis, Antineoplásicos, Herbal medicine