Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma

Autores
Laurent, Viviana Eunice; Otero, Laura L.; Vazquez, Valeria; Camarero, Sandra; Gabri, Mariano Rolando; Labrada, Maria; Garcia de Davila, Maria Teresa; Chantada, Guillermo Luis; Alonso, Daniel Fernando
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Extraocular dissemination is the main cause of death in patients with retinoblastoma in developing countries and there are few molecular markers that could be used for evaluation of minimal disseminated disease. The expression of the ganglioside GD2 is present in retinoblastoma cells metastatic to the bone marrow and the enzyme GD2 synthase activity is detected in neuroblastoma, which shares many phenotypic features with retinoblastoma. Our purpose was to optimize the detection of GD2 synthase expression by reverse transcription-polymerase chain reaction (RT-PCR) followed by nested-PCR in human retinoblastoma cell lines and patient samples. The optimization strategy was carried out by using the retinoblastoma cell lines Y79 and WERI-Rb1 and specific primers designed for the human sequence of the GD2 synthase mRNA. We detected GD2 synthase expression with at least 200 pg and 40 pg of total RNA extracted from cultured retinoblastoma cells, using a first round of RT-PCR amplification and a second round of nested-PCR, respectively. We have also confirmed the detection of GD2 synthase by RT-PCR and immunohistochemical expression of the ganglioside in human retinoblastoma tumors xenotransplanted in nude mice. In a study from tumor bank specimens from 8 retinoblastoma patients, we were able to demonstrate the presence of GD2 synthase mRNA in blood and cerebrospinal fluid samples in cases of extraocular dissemination of the tumor. The sequence was not detected in samples from children with low-risk disease or healthy adult volunteers. The detection of GD2 synthase mRNA through an optimized nested RT-PCR assay may be a promising tool for the assessment of minimal disseminated disease in enucleated patients.
Fil: Laurent, Viviana Eunice. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Servicio de Hemato-Oncología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina
Fil: Otero, Laura L.. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina
Fil: Vazquez, Valeria. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina
Fil: Camarero, Sandra. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; Argentina
Fil: Gabri, Mariano Rolando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina
Fil: Labrada, Maria. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; Argentina
Fil: Garcia de Davila, Maria Teresa. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; Argentina
Fil: Chantada, Guillermo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Servicio de Hemato-Oncología; Argentina
Fil: Alonso, Daniel Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina
Materia
GD2 synthase
Retinoblastoma
Molecular detection
RT-PCR
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/102452
Acceder
id CONICETDig_d032adf1ef4fcaacacd26e085ad64830
oai_identifier_str oai:ri.conicet.gov.ar:11336/102452
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Optimization of molecular detection of GD2 synthase mRNA in retinoblastomaLaurent, Viviana EuniceOtero, Laura L.Vazquez, ValeriaCamarero, SandraGabri, Mariano RolandoLabrada, MariaGarcia de Davila, Maria TeresaChantada, Guillermo LuisAlonso, Daniel FernandoGD2 synthaseRetinoblastomaMolecular detectionRT-PCRhttps://purl.org/becyt/ford/3.2https://purl.org/becyt/ford/3Extraocular dissemination is the main cause of death in patients with retinoblastoma in developing countries and there are few molecular markers that could be used for evaluation of minimal disseminated disease. The expression of the ganglioside GD2 is present in retinoblastoma cells metastatic to the bone marrow and the enzyme GD2 synthase activity is detected in neuroblastoma, which shares many phenotypic features with retinoblastoma. Our purpose was to optimize the detection of GD2 synthase expression by reverse transcription-polymerase chain reaction (RT-PCR) followed by nested-PCR in human retinoblastoma cell lines and patient samples. The optimization strategy was carried out by using the retinoblastoma cell lines Y79 and WERI-Rb1 and specific primers designed for the human sequence of the GD2 synthase mRNA. We detected GD2 synthase expression with at least 200 pg and 40 pg of total RNA extracted from cultured retinoblastoma cells, using a first round of RT-PCR amplification and a second round of nested-PCR, respectively. We have also confirmed the detection of GD2 synthase by RT-PCR and immunohistochemical expression of the ganglioside in human retinoblastoma tumors xenotransplanted in nude mice. In a study from tumor bank specimens from 8 retinoblastoma patients, we were able to demonstrate the presence of GD2 synthase mRNA in blood and cerebrospinal fluid samples in cases of extraocular dissemination of the tumor. The sequence was not detected in samples from children with low-risk disease or healthy adult volunteers. The detection of GD2 synthase mRNA through an optimized nested RT-PCR assay may be a promising tool for the assessment of minimal disseminated disease in enucleated patients.Fil: Laurent, Viviana Eunice. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Servicio de Hemato-Oncología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; ArgentinaFil: Otero, Laura L.. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; ArgentinaFil: Vazquez, Valeria. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; ArgentinaFil: Camarero, Sandra. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; ArgentinaFil: Gabri, Mariano Rolando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; ArgentinaFil: Labrada, Maria. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; ArgentinaFil: Garcia de Davila, Maria Teresa. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; ArgentinaFil: Chantada, Guillermo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Servicio de Hemato-Oncología; ArgentinaFil: Alonso, Daniel Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; ArgentinaSpandidos Publications2010-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/102452Laurent, Viviana Eunice; Otero, Laura L.; Vazquez, Valeria; Camarero, Sandra; Gabri, Mariano Rolando; et al.; Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma; Spandidos Publications; Molecular Medicine Reports; 3; 3-2010; 253-2591791-2997CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.spandidos-publications.com/mmr/3/2/253info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:01:35Zoai:ri.conicet.gov.ar:11336/102452instacron: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:34982025-10-22 11:01:35.835CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma
title Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma
spellingShingle Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma
Laurent, Viviana Eunice
GD2 synthase
Retinoblastoma
Molecular detection
RT-PCR
title_short Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma
title_full Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma
title_fullStr Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma
title_full_unstemmed Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma
title_sort Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma
dc.creator.none.fl_str_mv Laurent, Viviana Eunice
Otero, Laura L.
Vazquez, Valeria
Camarero, Sandra
Gabri, Mariano Rolando
Labrada, Maria
Garcia de Davila, Maria Teresa
Chantada, Guillermo Luis
Alonso, Daniel Fernando
author Laurent, Viviana Eunice
author_facet Laurent, Viviana Eunice
Otero, Laura L.
Vazquez, Valeria
Camarero, Sandra
Gabri, Mariano Rolando
Labrada, Maria
Garcia de Davila, Maria Teresa
Chantada, Guillermo Luis
Alonso, Daniel Fernando
author_role author
author2 Otero, Laura L.
Vazquez, Valeria
Camarero, Sandra
Gabri, Mariano Rolando
Labrada, Maria
Garcia de Davila, Maria Teresa
Chantada, Guillermo Luis
Alonso, Daniel Fernando
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv GD2 synthase
Retinoblastoma
Molecular detection
RT-PCR
topic GD2 synthase
Retinoblastoma
Molecular detection
RT-PCR
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.2
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Extraocular dissemination is the main cause of death in patients with retinoblastoma in developing countries and there are few molecular markers that could be used for evaluation of minimal disseminated disease. The expression of the ganglioside GD2 is present in retinoblastoma cells metastatic to the bone marrow and the enzyme GD2 synthase activity is detected in neuroblastoma, which shares many phenotypic features with retinoblastoma. Our purpose was to optimize the detection of GD2 synthase expression by reverse transcription-polymerase chain reaction (RT-PCR) followed by nested-PCR in human retinoblastoma cell lines and patient samples. The optimization strategy was carried out by using the retinoblastoma cell lines Y79 and WERI-Rb1 and specific primers designed for the human sequence of the GD2 synthase mRNA. We detected GD2 synthase expression with at least 200 pg and 40 pg of total RNA extracted from cultured retinoblastoma cells, using a first round of RT-PCR amplification and a second round of nested-PCR, respectively. We have also confirmed the detection of GD2 synthase by RT-PCR and immunohistochemical expression of the ganglioside in human retinoblastoma tumors xenotransplanted in nude mice. In a study from tumor bank specimens from 8 retinoblastoma patients, we were able to demonstrate the presence of GD2 synthase mRNA in blood and cerebrospinal fluid samples in cases of extraocular dissemination of the tumor. The sequence was not detected in samples from children with low-risk disease or healthy adult volunteers. The detection of GD2 synthase mRNA through an optimized nested RT-PCR assay may be a promising tool for the assessment of minimal disseminated disease in enucleated patients.
Fil: Laurent, Viviana Eunice. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Servicio de Hemato-Oncología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina
Fil: Otero, Laura L.. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina
Fil: Vazquez, Valeria. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina
Fil: Camarero, Sandra. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; Argentina
Fil: Gabri, Mariano Rolando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina
Fil: Labrada, Maria. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; Argentina
Fil: Garcia de Davila, Maria Teresa. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; Argentina
Fil: Chantada, Guillermo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Servicio de Hemato-Oncología; Argentina
Fil: Alonso, Daniel Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina
description Extraocular dissemination is the main cause of death in patients with retinoblastoma in developing countries and there are few molecular markers that could be used for evaluation of minimal disseminated disease. The expression of the ganglioside GD2 is present in retinoblastoma cells metastatic to the bone marrow and the enzyme GD2 synthase activity is detected in neuroblastoma, which shares many phenotypic features with retinoblastoma. Our purpose was to optimize the detection of GD2 synthase expression by reverse transcription-polymerase chain reaction (RT-PCR) followed by nested-PCR in human retinoblastoma cell lines and patient samples. The optimization strategy was carried out by using the retinoblastoma cell lines Y79 and WERI-Rb1 and specific primers designed for the human sequence of the GD2 synthase mRNA. We detected GD2 synthase expression with at least 200 pg and 40 pg of total RNA extracted from cultured retinoblastoma cells, using a first round of RT-PCR amplification and a second round of nested-PCR, respectively. We have also confirmed the detection of GD2 synthase by RT-PCR and immunohistochemical expression of the ganglioside in human retinoblastoma tumors xenotransplanted in nude mice. In a study from tumor bank specimens from 8 retinoblastoma patients, we were able to demonstrate the presence of GD2 synthase mRNA in blood and cerebrospinal fluid samples in cases of extraocular dissemination of the tumor. The sequence was not detected in samples from children with low-risk disease or healthy adult volunteers. The detection of GD2 synthase mRNA through an optimized nested RT-PCR assay may be a promising tool for the assessment of minimal disseminated disease in enucleated patients.
publishDate 2010
dc.date.none.fl_str_mv 2010-03
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/102452
Laurent, Viviana Eunice; Otero, Laura L.; Vazquez, Valeria; Camarero, Sandra; Gabri, Mariano Rolando; et al.; Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma; Spandidos Publications; Molecular Medicine Reports; 3; 3-2010; 253-259
1791-2997
CONICET Digital
CONICET
url http://hdl.handle.net/11336/102452
identifier_str_mv Laurent, Viviana Eunice; Otero, Laura L.; Vazquez, Valeria; Camarero, Sandra; Gabri, Mariano Rolando; et al.; Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma; Spandidos Publications; Molecular Medicine Reports; 3; 3-2010; 253-259
1791-2997
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.spandidos-publications.com/mmr/3/2/253
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Spandidos Publications
publisher.none.fl_str_mv Spandidos Publications
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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score 12.982451

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