Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7)

Autores
Chapana, Agostina Lucía; Carvelli, Flavia Lorena; Guarniolo, D.; Sosa, E.; Fernandez, R. A.; Barrera, Patricia Andrea; Sosa, Marcela; Caballero, P. A.
Año de publicación
2020
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Botulinum neurotoxin serotype A (BoNT A), produced by Clostridium botulinum, causes botulism and is used to treat multiple diseases. Its potential action in cancer therapy is currently being evaluated. In precedent studies, we have shown that BoNT A from native soil strains (Su), they have different characteristics to that of the prototype A Hall strain such as; greater specific toxic activity (AE) and differences in molecular structure and enzymatic activity against SNARE proteins. In this work, we study the action of an autochthonous BoNT A compared to the A Hall on tubulin in the MCF-7 cell line of breast carcinoma. The autochthonous BoNTs of the strain Su 1935 (Tupungato) and A Hall were used in their native form, purified by saline precipitation. The values of AE (LD50/mg protein) and electrophoretic characteristics under non-denaturing conditions were determined. MCF7 cells were cultured on coverslips and incubated with 250 and 500 LD50 of the BoNTs for 10, 25, 45, and 90 min. Later, the cells were fixed and processed for immunodetection. As primary antibodies were used anti-tubulin or anti-Golgina 97 and as secondary anti-mouse-Alexa 488. The preparations visualized by fluorescence microscopy. At 90 min incubation with 250 LD50, it was observed that ~90% of the cells were taken off and deformed by the action of the BoNT A 1935 and ~40% for the BoNT A Hall, while with 500 LD50 both toxins were deleterious to the cells. When the cells were incubated with the toxins for 25 min, a disruption of microtubules with both toxins was observed, the effect being greater with the BoNT A1935. This effect was accompanied by a redistribution of the Golgi apparatus. Western blotting showed the shape of new tubulin bands, possibly due to protein degradation. This effect was also greater in the BoNT 1935. These results show a cytotoxic action of BoNT A with disorganization of cell microtubules, being observed with greater intensity in the cells treated with the autochthonous BoNT A. The degradation of tubulin and its intracellular reorganization would be part of the deleterious action of this toxin on tumor cells, opening new perspectives for therapy against solid tumors.
Fil: Chapana, Agostina Lucía. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Carvelli, Flavia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Histología y Embriología D/mend Dr.m.burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Guarniolo, D.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina
Fil: Sosa, E.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina
Fil: Fernandez, R. A.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina
Fil: Barrera, Patricia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Histología y Embriología D/mend Dr.m.burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Sosa, Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Histología y Embriología D/mend Dr.m.burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Caballero, P. A.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina
XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo
San Luis
Argentina
Sociedad Biología de Cuyo
Materia
CLOSTRIDIUM BOTULINUM
BREAST CARCINOMA
BOTULINUM NEUROTOXIN
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/216285
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/216285
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7)Chapana, Agostina LucíaCarvelli, Flavia LorenaGuarniolo, D.Sosa, E.Fernandez, R. A.Barrera, Patricia AndreaSosa, MarcelaCaballero, P. A.CLOSTRIDIUM BOTULINUMBREAST CARCINOMABOTULINUM NEUROTOXINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Botulinum neurotoxin serotype A (BoNT A), produced by Clostridium botulinum, causes botulism and is used to treat multiple diseases. Its potential action in cancer therapy is currently being evaluated. In precedent studies, we have shown that BoNT A from native soil strains (Su), they have different characteristics to that of the prototype A Hall strain such as; greater specific toxic activity (AE) and differences in molecular structure and enzymatic activity against SNARE proteins. In this work, we study the action of an autochthonous BoNT A compared to the A Hall on tubulin in the MCF-7 cell line of breast carcinoma. The autochthonous BoNTs of the strain Su 1935 (Tupungato) and A Hall were used in their native form, purified by saline precipitation. The values of AE (LD50/mg protein) and electrophoretic characteristics under non-denaturing conditions were determined. MCF7 cells were cultured on coverslips and incubated with 250 and 500 LD50 of the BoNTs for 10, 25, 45, and 90 min. Later, the cells were fixed and processed for immunodetection. As primary antibodies were used anti-tubulin or anti-Golgina 97 and as secondary anti-mouse-Alexa 488. The preparations visualized by fluorescence microscopy. At 90 min incubation with 250 LD50, it was observed that ~90% of the cells were taken off and deformed by the action of the BoNT A 1935 and ~40% for the BoNT A Hall, while with 500 LD50 both toxins were deleterious to the cells. When the cells were incubated with the toxins for 25 min, a disruption of microtubules with both toxins was observed, the effect being greater with the BoNT A1935. This effect was accompanied by a redistribution of the Golgi apparatus. Western blotting showed the shape of new tubulin bands, possibly due to protein degradation. This effect was also greater in the BoNT 1935. These results show a cytotoxic action of BoNT A with disorganization of cell microtubules, being observed with greater intensity in the cells treated with the autochthonous BoNT A. The degradation of tubulin and its intracellular reorganization would be part of the deleterious action of this toxin on tumor cells, opening new perspectives for therapy against solid tumors.Fil: Chapana, Agostina Lucía. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Carvelli, Flavia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Histología y Embriología D/mend Dr.m.burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Guarniolo, D.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; ArgentinaFil: Sosa, E.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; ArgentinaFil: Fernandez, R. A.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; ArgentinaFil: Barrera, Patricia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Histología y Embriología D/mend Dr.m.burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Sosa, Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Histología y Embriología D/mend Dr.m.burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Caballero, P. A.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; ArgentinaXXXVII Reunión Científica Anual de la Sociedad de Biología de CuyoSan LuisArgentinaSociedad Biología de CuyoTech Science Press2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/216285Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7); XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo; San Luis; Argentina; 2019; 12-130327-9545CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://sbcuyo.org.ar/wp-content/uploads/2019/12/Libro-BIOCELL-2019.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:59:29Zoai:ri.conicet.gov.ar:11336/216285instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:59:29.425CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7)
title Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7)
spellingShingle Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7)
Chapana, Agostina Lucía
CLOSTRIDIUM BOTULINUM
BREAST CARCINOMA
BOTULINUM NEUROTOXIN
title_short Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7)
title_full Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7)
title_fullStr Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7)
title_full_unstemmed Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7)
title_sort Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7)
dc.creator.none.fl_str_mv Chapana, Agostina Lucía
Carvelli, Flavia Lorena
Guarniolo, D.
Sosa, E.
Fernandez, R. A.
Barrera, Patricia Andrea
Sosa, Marcela
Caballero, P. A.
author Chapana, Agostina Lucía
author_facet Chapana, Agostina Lucía
Carvelli, Flavia Lorena
Guarniolo, D.
Sosa, E.
Fernandez, R. A.
Barrera, Patricia Andrea
Sosa, Marcela
Caballero, P. A.
author_role author
author2 Carvelli, Flavia Lorena
Guarniolo, D.
Sosa, E.
Fernandez, R. A.
Barrera, Patricia Andrea
Sosa, Marcela
Caballero, P. A.
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CLOSTRIDIUM BOTULINUM
BREAST CARCINOMA
BOTULINUM NEUROTOXIN
topic CLOSTRIDIUM BOTULINUM
BREAST CARCINOMA
BOTULINUM NEUROTOXIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Botulinum neurotoxin serotype A (BoNT A), produced by Clostridium botulinum, causes botulism and is used to treat multiple diseases. Its potential action in cancer therapy is currently being evaluated. In precedent studies, we have shown that BoNT A from native soil strains (Su), they have different characteristics to that of the prototype A Hall strain such as; greater specific toxic activity (AE) and differences in molecular structure and enzymatic activity against SNARE proteins. In this work, we study the action of an autochthonous BoNT A compared to the A Hall on tubulin in the MCF-7 cell line of breast carcinoma. The autochthonous BoNTs of the strain Su 1935 (Tupungato) and A Hall were used in their native form, purified by saline precipitation. The values of AE (LD50/mg protein) and electrophoretic characteristics under non-denaturing conditions were determined. MCF7 cells were cultured on coverslips and incubated with 250 and 500 LD50 of the BoNTs for 10, 25, 45, and 90 min. Later, the cells were fixed and processed for immunodetection. As primary antibodies were used anti-tubulin or anti-Golgina 97 and as secondary anti-mouse-Alexa 488. The preparations visualized by fluorescence microscopy. At 90 min incubation with 250 LD50, it was observed that ~90% of the cells were taken off and deformed by the action of the BoNT A 1935 and ~40% for the BoNT A Hall, while with 500 LD50 both toxins were deleterious to the cells. When the cells were incubated with the toxins for 25 min, a disruption of microtubules with both toxins was observed, the effect being greater with the BoNT A1935. This effect was accompanied by a redistribution of the Golgi apparatus. Western blotting showed the shape of new tubulin bands, possibly due to protein degradation. This effect was also greater in the BoNT 1935. These results show a cytotoxic action of BoNT A with disorganization of cell microtubules, being observed with greater intensity in the cells treated with the autochthonous BoNT A. The degradation of tubulin and its intracellular reorganization would be part of the deleterious action of this toxin on tumor cells, opening new perspectives for therapy against solid tumors.
Fil: Chapana, Agostina Lucía. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Carvelli, Flavia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Histología y Embriología D/mend Dr.m.burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Guarniolo, D.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina
Fil: Sosa, E.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina
Fil: Fernandez, R. A.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina
Fil: Barrera, Patricia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Histología y Embriología D/mend Dr.m.burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Sosa, Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Histología y Embriología D/mend Dr.m.burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Caballero, P. A.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina
XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo
San Luis
Argentina
Sociedad Biología de Cuyo
description Botulinum neurotoxin serotype A (BoNT A), produced by Clostridium botulinum, causes botulism and is used to treat multiple diseases. Its potential action in cancer therapy is currently being evaluated. In precedent studies, we have shown that BoNT A from native soil strains (Su), they have different characteristics to that of the prototype A Hall strain such as; greater specific toxic activity (AE) and differences in molecular structure and enzymatic activity against SNARE proteins. In this work, we study the action of an autochthonous BoNT A compared to the A Hall on tubulin in the MCF-7 cell line of breast carcinoma. The autochthonous BoNTs of the strain Su 1935 (Tupungato) and A Hall were used in their native form, purified by saline precipitation. The values of AE (LD50/mg protein) and electrophoretic characteristics under non-denaturing conditions were determined. MCF7 cells were cultured on coverslips and incubated with 250 and 500 LD50 of the BoNTs for 10, 25, 45, and 90 min. Later, the cells were fixed and processed for immunodetection. As primary antibodies were used anti-tubulin or anti-Golgina 97 and as secondary anti-mouse-Alexa 488. The preparations visualized by fluorescence microscopy. At 90 min incubation with 250 LD50, it was observed that ~90% of the cells were taken off and deformed by the action of the BoNT A 1935 and ~40% for the BoNT A Hall, while with 500 LD50 both toxins were deleterious to the cells. When the cells were incubated with the toxins for 25 min, a disruption of microtubules with both toxins was observed, the effect being greater with the BoNT A1935. This effect was accompanied by a redistribution of the Golgi apparatus. Western blotting showed the shape of new tubulin bands, possibly due to protein degradation. This effect was also greater in the BoNT 1935. These results show a cytotoxic action of BoNT A with disorganization of cell microtubules, being observed with greater intensity in the cells treated with the autochthonous BoNT A. The degradation of tubulin and its intracellular reorganization would be part of the deleterious action of this toxin on tumor cells, opening new perspectives for therapy against solid tumors.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/216285
Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7); XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo; San Luis; Argentina; 2019; 12-13
0327-9545
CONICET Digital
CONICET
url http://hdl.handle.net/11336/216285
identifier_str_mv Effect of botulinum neurotoxin of Mendoza clostridium botulinum atrain on tubulin in breast carcinoma cells (MCF 7); XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo; San Luis; Argentina; 2019; 12-13
0327-9545
CONICET Digital
CONICET
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language eng
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rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
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dc.publisher.none.fl_str_mv Tech Science Press
publisher.none.fl_str_mv Tech Science Press
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