Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells

Autores
Chapana, Agostina Lucía; Guarniolo, D.; Carvelli, Flavia Lorena; Sosa, E.; Fernández, R. A.; Sosa, M. A.; Caballero, P. A.
Año de publicación
2021
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The botulinum neurotoxin serotype A (BoNT A) produced by Clostridium botulinum, which causes botulism, is used for the treatment of multiple neurological diseases and its therapeutic action against cancer is currently being evaluated. In previous studies, we have shown that BoNT A from autochthonous soil strains (Su) have different properties than the reference A Hall strain. Among these, its molecular structure, its enzymatic activity against brain SNARE proteins and its greater specific toxic activity (AE) stand out. In cells from human mammary carcinoma (MCF-7) treated with BoNTs for 45 min, we found a marked effect on the expression of cytoskeletal proteins. Therefore, in this work, we delve into the study of the action of autochthonous BoNTs A and prototype A Hall on the distribution of actin and tubulin in these cells. Native forms of autochthonous BoNT (Su strains 1935 and 1891, Tupungato) and prototype A Hall were purified by saline precipitation. Their AE values (LD50 / mg protein) were established and their electrophoretic characteristics were evaluated under non-denaturing conditions. 250 LD50 of the BoNTs were incubated to MCF-7 cell cultures for 10 or 25 min. Later, the cells were fixed and processed for indirect immunofluorescence with the use of specific antibodies that recognize tubulin or actin. The samples were visualized by fluorescence microscopy. At the two times evaluated, the three types of BoNTs produced a marked redistribution of the actin cytoskeleton, patch form, on areas coinciding with the plasma membrane. Tubulin was redistributed to multiple areas with high signal density at 10 min of incubation only in the presence of BoNT 1891. At 25 min of incubation, the cells treated with BoNTs 1891 and 1935 showed this effect, while in those incubated with A Hall, the distribution of these proteins was not modified. The notable alterations in the distribution of components of the tumor cell cytoskeleton by BoNT from native strains of Mendoza soils open new perspectives for therapy against solid tumors.
Fil: Chapana, Agostina Lucía. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Guarniolo, D.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Carvelli, Flavia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Sosa, E.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Fernández, R. A.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Sosa, M. A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Caballero, P. A.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
IV Reunión Conjunta de Sociedades de Biología de la República Argentina
Argentina
Sociedad de Biología de Cuyo
Sociedad Argentina de Biología
Sociedad de Biología de Rosario
Sociedad Chilena de Reproducción y Desarrollo
Asociación de Biología de Tucumán
Sociedad de Biología de Córdoba
Materia
BOTULINUM NEUROTOXIN
BREAST CANCER
CYTOSKELETON
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:11336/196118
Acceder
id CONICETDig_3b966a50ce753951af37bab9f61dc8f9
oai_identifier_str oai:ri.conicet.gov.ar:11336/196118
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cellsChapana, Agostina LucíaGuarniolo, D.Carvelli, Flavia LorenaSosa, E.Fernández, R. A.Sosa, M. A.Caballero, P. A.BOTULINUM NEUROTOXINBREAST CANCERCYTOSKELETONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The botulinum neurotoxin serotype A (BoNT A) produced by Clostridium botulinum, which causes botulism, is used for the treatment of multiple neurological diseases and its therapeutic action against cancer is currently being evaluated. In previous studies, we have shown that BoNT A from autochthonous soil strains (Su) have different properties than the reference A Hall strain. Among these, its molecular structure, its enzymatic activity against brain SNARE proteins and its greater specific toxic activity (AE) stand out. In cells from human mammary carcinoma (MCF-7) treated with BoNTs for 45 min, we found a marked effect on the expression of cytoskeletal proteins. Therefore, in this work, we delve into the study of the action of autochthonous BoNTs A and prototype A Hall on the distribution of actin and tubulin in these cells. Native forms of autochthonous BoNT (Su strains 1935 and 1891, Tupungato) and prototype A Hall were purified by saline precipitation. Their AE values (LD50 / mg protein) were established and their electrophoretic characteristics were evaluated under non-denaturing conditions. 250 LD50 of the BoNTs were incubated to MCF-7 cell cultures for 10 or 25 min. Later, the cells were fixed and processed for indirect immunofluorescence with the use of specific antibodies that recognize tubulin or actin. The samples were visualized by fluorescence microscopy. At the two times evaluated, the three types of BoNTs produced a marked redistribution of the actin cytoskeleton, patch form, on areas coinciding with the plasma membrane. Tubulin was redistributed to multiple areas with high signal density at 10 min of incubation only in the presence of BoNT 1891. At 25 min of incubation, the cells treated with BoNTs 1891 and 1935 showed this effect, while in those incubated with A Hall, the distribution of these proteins was not modified. The notable alterations in the distribution of components of the tumor cell cytoskeleton by BoNT from native strains of Mendoza soils open new perspectives for therapy against solid tumors.Fil: Chapana, Agostina Lucía. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Guarniolo, D.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; ArgentinaFil: Carvelli, Flavia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Sosa, E.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; ArgentinaFil: Fernández, R. A.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; ArgentinaFil: Sosa, M. A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Caballero, P. A.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; ArgentinaIV Reunión Conjunta de Sociedades de Biología de la República ArgentinaArgentinaSociedad de Biología de CuyoSociedad Argentina de BiologíaSociedad de Biología de RosarioSociedad Chilena de Reproducción y DesarrolloAsociación de Biología de TucumánSociedad de Biología de CórdobaTech Science PressPiezzi, Ramon Salvador2021info: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/11336/196118Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells; IV Reunión Conjunta de Sociedades de Biología de la República Argentina; Argentina; 2020; 16-160327-95451667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.techscience.com/uploads/attached/file/20210806/20210806053124_35488.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:54:40Zoai:ri.conicet.gov.ar:11336/196118instacron: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 09:54:40.593CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells
title Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells
spellingShingle Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells
Chapana, Agostina Lucía
BOTULINUM NEUROTOXIN
BREAST CANCER
CYTOSKELETON
title_short Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells
title_full Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells
title_fullStr Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells
title_full_unstemmed Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells
title_sort Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells
dc.creator.none.fl_str_mv Chapana, Agostina Lucía
Guarniolo, D.
Carvelli, Flavia Lorena
Sosa, E.
Fernández, R. A.
Sosa, M. A.
Caballero, P. A.
author Chapana, Agostina Lucía
author_facet Chapana, Agostina Lucía
Guarniolo, D.
Carvelli, Flavia Lorena
Sosa, E.
Fernández, R. A.
Sosa, M. A.
Caballero, P. A.
author_role author
author2 Guarniolo, D.
Carvelli, Flavia Lorena
Sosa, E.
Fernández, R. A.
Sosa, M. A.
Caballero, P. A.
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Piezzi, Ramon Salvador
dc.subject.none.fl_str_mv BOTULINUM NEUROTOXIN
BREAST CANCER
CYTOSKELETON
topic BOTULINUM NEUROTOXIN
BREAST CANCER
CYTOSKELETON
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The botulinum neurotoxin serotype A (BoNT A) produced by Clostridium botulinum, which causes botulism, is used for the treatment of multiple neurological diseases and its therapeutic action against cancer is currently being evaluated. In previous studies, we have shown that BoNT A from autochthonous soil strains (Su) have different properties than the reference A Hall strain. Among these, its molecular structure, its enzymatic activity against brain SNARE proteins and its greater specific toxic activity (AE) stand out. In cells from human mammary carcinoma (MCF-7) treated with BoNTs for 45 min, we found a marked effect on the expression of cytoskeletal proteins. Therefore, in this work, we delve into the study of the action of autochthonous BoNTs A and prototype A Hall on the distribution of actin and tubulin in these cells. Native forms of autochthonous BoNT (Su strains 1935 and 1891, Tupungato) and prototype A Hall were purified by saline precipitation. Their AE values (LD50 / mg protein) were established and their electrophoretic characteristics were evaluated under non-denaturing conditions. 250 LD50 of the BoNTs were incubated to MCF-7 cell cultures for 10 or 25 min. Later, the cells were fixed and processed for indirect immunofluorescence with the use of specific antibodies that recognize tubulin or actin. The samples were visualized by fluorescence microscopy. At the two times evaluated, the three types of BoNTs produced a marked redistribution of the actin cytoskeleton, patch form, on areas coinciding with the plasma membrane. Tubulin was redistributed to multiple areas with high signal density at 10 min of incubation only in the presence of BoNT 1891. At 25 min of incubation, the cells treated with BoNTs 1891 and 1935 showed this effect, while in those incubated with A Hall, the distribution of these proteins was not modified. The notable alterations in the distribution of components of the tumor cell cytoskeleton by BoNT from native strains of Mendoza soils open new perspectives for therapy against solid tumors.
Fil: Chapana, Agostina Lucía. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Guarniolo, D.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Carvelli, Flavia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Sosa, E.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Fernández, R. A.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Sosa, M. A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Caballero, P. A.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
IV Reunión Conjunta de Sociedades de Biología de la República Argentina
Argentina
Sociedad de Biología de Cuyo
Sociedad Argentina de Biología
Sociedad de Biología de Rosario
Sociedad Chilena de Reproducción y Desarrollo
Asociación de Biología de Tucumán
Sociedad de Biología de Córdoba
description The botulinum neurotoxin serotype A (BoNT A) produced by Clostridium botulinum, which causes botulism, is used for the treatment of multiple neurological diseases and its therapeutic action against cancer is currently being evaluated. In previous studies, we have shown that BoNT A from autochthonous soil strains (Su) have different properties than the reference A Hall strain. Among these, its molecular structure, its enzymatic activity against brain SNARE proteins and its greater specific toxic activity (AE) stand out. In cells from human mammary carcinoma (MCF-7) treated with BoNTs for 45 min, we found a marked effect on the expression of cytoskeletal proteins. Therefore, in this work, we delve into the study of the action of autochthonous BoNTs A and prototype A Hall on the distribution of actin and tubulin in these cells. Native forms of autochthonous BoNT (Su strains 1935 and 1891, Tupungato) and prototype A Hall were purified by saline precipitation. Their AE values (LD50 / mg protein) were established and their electrophoretic characteristics were evaluated under non-denaturing conditions. 250 LD50 of the BoNTs were incubated to MCF-7 cell cultures for 10 or 25 min. Later, the cells were fixed and processed for indirect immunofluorescence with the use of specific antibodies that recognize tubulin or actin. The samples were visualized by fluorescence microscopy. At the two times evaluated, the three types of BoNTs produced a marked redistribution of the actin cytoskeleton, patch form, on areas coinciding with the plasma membrane. Tubulin was redistributed to multiple areas with high signal density at 10 min of incubation only in the presence of BoNT 1891. At 25 min of incubation, the cells treated with BoNTs 1891 and 1935 showed this effect, while in those incubated with A Hall, the distribution of these proteins was not modified. The notable alterations in the distribution of components of the tumor cell cytoskeleton by BoNT from native strains of Mendoza soils open new perspectives for therapy against solid tumors.
publishDate 2021
dc.date.none.fl_str_mv 2021
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/11336/196118
Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells; IV Reunión Conjunta de Sociedades de Biología de la República Argentina; Argentina; 2020; 16-16
0327-9545
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/196118
identifier_str_mv Effect of botulinum neurotoxins from Mendoza of clostridium botulinum strains on cytoskeletal proteins of mammary tumor cells; IV Reunión Conjunta de Sociedades de Biología de la República Argentina; Argentina; 2020; 16-16
0327-9545
1667-5746
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.techscience.com/uploads/attached/file/20210806/20210806053124_35488.pdf
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Tech Science Press
publisher.none.fl_str_mv Tech Science Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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