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

Abstract

BackgroundMyeloproliferative 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.ObjectiveTo evaluate the effects of l-amino acid oxidase from C. rhodostoma snake venom in the apoptotic machinery of JAK2-mutated cell lines.MethodsThe 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.Resultsl-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.Conclusionsl-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.

Cytotoxic, Antiproliferative and Apoptosis‐inducing Activity of L‐Amino Acid Oxidase from Malaysian Calloselasma rhodostoma on Human Colon Cancer Cells

“…The present findings are in agreement with several LAAO studies from various snake species induced apoptosis in human cancer cell lines such as HeLa [21,45], MM6 [46], Jurkat cells [47] and RBR 17T (human malignant glioma cells) [48]. Moreover, the activation of caspase enzymes, such as caspase-3, was attributed to cancer cell lines, such as SET-2 and CML cells when exposed to CR-LAAO [30,31]. Besides caspase-3, Bcl-2 is an important indicator of apoptosis induction.…”

“…CR‐LAAO demonstrated cytotoxic activity against yeast, HL‐60 (human promyelocytic leukaemia cells) and HepG2 (hepatocarcinoma) cell lines . Moreover, the toxin was capable of inducing apoptosis in HEL 92.1.7 (human bone marrow erythrol), SET‐2 (human megakaryoblastic leukaemic cells) and CML (chronic myeloid leukaemia) cells . However, cytotoxic and apoptotic activities of CR‐LAAO in human colon cancer cells have not been reported.…”

Bothrops jararacaandBothrops erythromelasSnake Venoms Promote Cell Cycle Arrest and Induce Apoptosis via the Mitochondrial Depolarization of Cervical Cancer Cells

“…Recently, Tavares et al (2016) observed the induction of cytotoxicity by L-amino acid oxidase activity from the venom of Calloselasma rhodostoma species in bone marrow erythroleukemia cell lines (HEL92.1.7 cells) and megakaryoblastic leukemia (Set-2 cells) [27]. Other studies with adenocarcinoma cell lines of human colon LS174 (p53wt), HCT116 (p53wt), and HT29 (p53mut) found that Lebein (disintegrin heterodimeric) isolated from the venom of Macrovipera lebetina snake significantly inhibited cell viability [28].…”

BthTX-I from Bothrops jararacussu induces apoptosis in human breast cancer cell lines and decreases cancer stem cell subpopulation

“…Furthermore, BpirLAAO-I induced apoptosis and potentiated the tyronise kinase inhibitor effect on BCR-ABL + cells. Additionally, Tavares et al [8] reported an L-amino-acid oxidase from C. rhodostoma (CR-LAAO) snake venom as a potential antineoplastic agent against HEL 92.1.7 and SET-2 JAK2V617F cell lines derived from myeloproliferative neoplasm patients. Moreover, the cytotoxins CT1 and CT2 from Naja oxiana , CT3 from Naja kaouthia and CT1 from Naja haje showed an important cytotoxicity, mainly mediated by lysosome rupture, against lung adenocarcinoma A549 and promyelocytic leukemia HL60 cells [9].…”