l-Amino acid oxidase isolated from Calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in JAK2V617F-positive cell lines

Autor(es)

Cristiane Tavares ^[1] ; Thaís Maciel ^[2] ; Sandra Burin ^[3] ; Luciana Ambrósio ^[4] ; Sandro Ghisla ^[5] ; Suely Sampaio ^[6] ; Fabíola Castro ^[7]

Afiliação do(s) autor(es)

^[1] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Brasil

^[2] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Brasil

^[3] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Brasil

^[4] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Brasil

^[5] Universität Konstanz - Alemanha

^[6] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Brasil

^[7] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Brasil

• Tipo de documento: Artigo Científico
• Fonte: Revista Brasileira de Hematologia e Hemoterapia; v. 38, n. 2, p. 128-134, 2016-06-00.

Resumo

ABSTRACT BACKGROUND: Myeloproliferative neoplasms are Philadelphia chromosome-negative diseases characterized by hyperproliferation of mature myeloid cells, associated or not with the Janus kinase 2 tyrosine kinase mutation, JAK2V617F. As there is no curative therapy, researchers have been investigating new drugs to treat myeloproliferative neoplasms, including l-amino acid oxidase from Calloselasma rhodostoma snake venom (CR-LAAO), which is a toxin capable of eliciting apoptosis in several tumor cell lines. OBJECTIVE: To evaluate the effects of l-amino acid oxidase from C. rhodostoma snake venom in the apoptotic machinery of JAK2-mutated cell lines. METHODS: The HEL 92.1.7 and SET-2 cell lines were cultured with l-amino acid oxidase and catalase for 12 h at 37 °C in 5% carbon dioxide. The cell viability was assessed by the multi-table tournament method, the level of apoptosis was measured by flow cytometry, and the expression of cysteine-dependent aspartate-specific proteases and cleaved Poly(ADP-ribose) polymerase were analyzed by Western blotting. RESULTS: l-Amino acid oxidase from C. rhodostoma snake venom was cytotoxic to HEL 92.1.7 and SET-2 cells (50% inhibitory concentration = 0.15 µg/mL and 1.5 µg/mL, respectively) and induced apoptosis in a concentration-dependent manner. Cell treatment with catalase mitigated the l-amino acid oxidase toxicity, indicating that hydrogen peroxide is a key component of its cytotoxic effect.The activated caspases 3 and 8 expression and cleaved PARP in HEL 92.1.7 and SET-2 cells confirmed the apoptosis activation by CR-LAAO. CONCLUSIONS: l-Amino acid oxidase from C. rhodostoma snake venom is a potential antineoplastic agent against HEL 92.1.7 and SET-2 JAK2V617F-positive cells as it activates the extrinsic apoptosis pathway. (AU)

• Processo FAPESP: 11/23236-4 - Toxinas animais nativas e recombinantes: análise funcional, estrutural e molecular
• Beneficiário: Suely Vilela
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 14/19127-3 - Efeito da LAAO de Calloselasma rhodostoma e Bothrops moojeni na maquinaria apoptótica de linhagens JAK2V617F positivas de neoplasias mieloproliferativas
• Beneficiário: Thaís Fontanezi Maciel
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica