l-Amino acid oxidase isolated from Calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in JAK2V617F-positive cell lines
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 investi...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2016-06-01
|
| Series: | Revista Brasileira de Hematologia e Hemoterapia |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-84842016000200128&lng=en&tlng=en |
| _version_ | 1818572064666157056 |
|---|---|
| author | Cristiane Tavares Thaís Maciel Sandra Burin Luciana Ambrósio Sandro Ghisla Suely Sampaio Fabíola Castro |
| author_facet | Cristiane Tavares Thaís Maciel Sandra Burin Luciana Ambrósio Sandro Ghisla Suely Sampaio Fabíola Castro |
| author_sort | Cristiane Tavares |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-14T18:52:26Z |
| format | Article |
| id | doaj.art-445b3d9503574caf9fd7d66127a27d07 |
| institution | Directory Open Access Journal |
| issn | 1806-0870 |
| language | English |
| last_indexed | 2024-12-14T18:52:26Z |
| publishDate | 2016-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Revista Brasileira de Hematologia e Hemoterapia |
| spelling | doaj.art-445b3d9503574caf9fd7d66127a27d072022-12-21T22:51:11ZengElsevierRevista Brasileira de Hematologia e Hemoterapia1806-08702016-06-0138212813410.1016/j.bjhh.2016.03.004S1516-84842016000200128l-Amino acid oxidase isolated from Calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in JAK2V617F-positive cell linesCristiane TavaresThaís MacielSandra BurinLuciana AmbrósioSandro GhislaSuely SampaioFabíola CastroABSTRACT 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-84842016000200128&lng=en&tlng=enMyeloproliferative neoplasmsApoptosisl-Amino acid oxidaseCalloselasma rhodostomaJanus kinase 2 mutation |
| spellingShingle | Cristiane Tavares Thaís Maciel Sandra Burin Luciana Ambrósio Sandro Ghisla Suely Sampaio Fabíola Castro l-Amino acid oxidase isolated from Calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in JAK2V617F-positive cell lines Revista Brasileira de Hematologia e Hemoterapia Myeloproliferative neoplasms Apoptosis l-Amino acid oxidase Calloselasma rhodostoma Janus kinase 2 mutation |
| title | l-Amino acid oxidase isolated from Calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in JAK2V617F-positive cell lines |
| title_full | l-Amino acid oxidase isolated from Calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in JAK2V617F-positive cell lines |
| title_fullStr | l-Amino acid oxidase isolated from Calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in JAK2V617F-positive cell lines |
| title_full_unstemmed | l-Amino acid oxidase isolated from Calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in JAK2V617F-positive cell lines |
| title_short | l-Amino acid oxidase isolated from Calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in JAK2V617F-positive cell lines |
| title_sort | l amino acid oxidase isolated from calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in jak2v617f positive cell lines |
| topic | Myeloproliferative neoplasms Apoptosis l-Amino acid oxidase Calloselasma rhodostoma Janus kinase 2 mutation |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-84842016000200128&lng=en&tlng=en |
| work_keys_str_mv | AT cristianetavares laminoacidoxidaseisolatedfromcalloselasmarhodostomasnakevenominducescytotoxicityandapoptosisinjak2v617fpositivecelllines AT thaismaciel laminoacidoxidaseisolatedfromcalloselasmarhodostomasnakevenominducescytotoxicityandapoptosisinjak2v617fpositivecelllines AT sandraburin laminoacidoxidaseisolatedfromcalloselasmarhodostomasnakevenominducescytotoxicityandapoptosisinjak2v617fpositivecelllines AT lucianaambrosio laminoacidoxidaseisolatedfromcalloselasmarhodostomasnakevenominducescytotoxicityandapoptosisinjak2v617fpositivecelllines AT sandroghisla laminoacidoxidaseisolatedfromcalloselasmarhodostomasnakevenominducescytotoxicityandapoptosisinjak2v617fpositivecelllines AT suelysampaio laminoacidoxidaseisolatedfromcalloselasmarhodostomasnakevenominducescytotoxicityandapoptosisinjak2v617fpositivecelllines AT fabiolacastro laminoacidoxidaseisolatedfromcalloselasmarhodostomasnakevenominducescytotoxicityandapoptosisinjak2v617fpositivecelllines |