Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids

Autores
Finocchiaro, L.M.E.; Bumaschny, V.F.; Karara, A.L.; Fiszman, G.L.; Casais, C.C.; Glikin, G.C.
Año de publicación
2004
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
We have developed multicellular spheroids (MCS) established from LM05e and LM3 spontaneous Balb/c-murine mammary adenocarcinoma and B16 C57-murine melanoma derived cell lines as an in vitro model to study the efficacy of the herpes simplex virus thymidine kinase/ganciclovir (HSVtk/GCV) suicide system. We demonstrated for the first time that HSVtk-expressing cells assembled as MCS manifested a GCV resistance phenotype compared to the same cells grown as sparse monolayers. HSVtk-expressing LM05e, LM3 and B16 spheroids were 16-, three- and nine-fold less sensitive to GCV than their respective monolayers, even though they could express transgenes 10-, eight- and five-fold more efficiently. Mixed populations of HSVtk- and their respective βgal-expressing cells displayed a cell-type specific bystander effect that was higher in monolayers than in MCS. However, HSVtk-expressing cells in two- or three-dimensional cultures were always significantly more sensitive to GCV than the βgal-expressing counterparts, supporting the feasibility of this suicide approach in vivo. We present evidence showing that HSVtk-expressing tumor cells, when transferred from monolayers to MCS, displayed: (i) lower GCV cytotoxic activity and bystander effect; (ii) higher and efficient expression of genes transferred as lipoplexes; (iii) lower cell proliferation rates; and (iv) changes in intracellular Bax/Bcl-xL rheostat of mitochondria-mediated apoptosis.
Fil:Finocchiaro, L.M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Casais, C.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Glikin, G.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fuente
Cancer Gene Ther. 2004;11(5):333-345
Materia
Lipofection
Murine adenocarcinoma
Suicide gene therapy
ganciclovir
suicide substrate
thymidine kinase
animal cell
animal tissue
apoptosis
article
breast adenocarcinoma
bystander effect
cell growth
cell proliferation
cell specificity
cell type
controlled study
drug cytotoxicity
drug efficacy
gene expression regulation
gene transfer
Herpes simplex virus
in vitro study
in vivo study
intracellular transport
melanoma cell
mitochondrion
monolayer culture
mouse
multicellular spheroid
nonhuman
phenotype
population structure
priority journal
protein assembly
sensitivity analysis
statistical significance
transgene
virus resistance
Adenocarcinoma
Animals
Antiviral Agents
bcl-2-Associated X Protein
beta-Galactosidase
Bystander Effect
Cell Division
Cell Line, Tumor
Drug Resistance
Ganciclovir
Gene Therapy
Mammary Neoplasms, Animal
Melanoma
Mice
Mice, Inbred BALB C
Proto-Oncogene Proteins c-bcl-2
Simplexvirus
Spheroids, Cellular
Thymidine Kinase
Transfection
Viral Proteins
Animalia
Murinae
Simplexvirus
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by/2.5/ar
Repositorio
Biblioteca Digital (UBA-FCEN)
Institución
Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
OAI Identificador
paperaa:paper_09291903_v11_n5_p333_Finocchiaro

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oai_identifier_str paperaa:paper_09291903_v11_n5_p333_Finocchiaro
network_acronym_str BDUBAFCEN
repository_id_str 1896
network_name_str Biblioteca Digital (UBA-FCEN)
spelling Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroidsFinocchiaro, L.M.E.Bumaschny, V.F.Karara, A.L.Fiszman, G.L.Casais, C.C.Glikin, G.C.LipofectionMurine adenocarcinomaSuicide gene therapyganciclovirsuicide substratethymidine kinaseanimal cellanimal tissueapoptosisarticlebreast adenocarcinomabystander effectcell growthcell proliferationcell specificitycell typecontrolled studydrug cytotoxicitydrug efficacygene expression regulationgene transferHerpes simplex virusin vitro studyin vivo studyintracellular transportmelanoma cellmitochondrionmonolayer culturemousemulticellular spheroidnonhumanphenotypepopulation structurepriority journalprotein assemblysensitivity analysisstatistical significancetransgenevirus resistanceAdenocarcinomaAnimalsAntiviral Agentsbcl-2-Associated X Proteinbeta-GalactosidaseBystander EffectCell DivisionCell Line, TumorDrug ResistanceGanciclovirGene TherapyMammary Neoplasms, AnimalMelanomaMiceMice, Inbred BALB CProto-Oncogene Proteins c-bcl-2SimplexvirusSpheroids, CellularThymidine KinaseTransfectionViral ProteinsAnimaliaMurinaeSimplexvirusWe have developed multicellular spheroids (MCS) established from LM05e and LM3 spontaneous Balb/c-murine mammary adenocarcinoma and B16 C57-murine melanoma derived cell lines as an in vitro model to study the efficacy of the herpes simplex virus thymidine kinase/ganciclovir (HSVtk/GCV) suicide system. We demonstrated for the first time that HSVtk-expressing cells assembled as MCS manifested a GCV resistance phenotype compared to the same cells grown as sparse monolayers. HSVtk-expressing LM05e, LM3 and B16 spheroids were 16-, three- and nine-fold less sensitive to GCV than their respective monolayers, even though they could express transgenes 10-, eight- and five-fold more efficiently. Mixed populations of HSVtk- and their respective βgal-expressing cells displayed a cell-type specific bystander effect that was higher in monolayers than in MCS. However, HSVtk-expressing cells in two- or three-dimensional cultures were always significantly more sensitive to GCV than the βgal-expressing counterparts, supporting the feasibility of this suicide approach in vivo. We present evidence showing that HSVtk-expressing tumor cells, when transferred from monolayers to MCS, displayed: (i) lower GCV cytotoxic activity and bystander effect; (ii) higher and efficient expression of genes transferred as lipoplexes; (iii) lower cell proliferation rates; and (iv) changes in intracellular Bax/Bcl-xL rheostat of mitochondria-mediated apoptosis.Fil:Finocchiaro, L.M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Casais, C.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Glikin, G.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.2004info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12110/paper_09291903_v11_n5_p333_FinocchiaroCancer Gene Ther. 2004;11(5):333-345reponame:Biblioteca Digital (UBA-FCEN)instname:Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturalesinstacron:UBA-FCENenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/2.5/ar2025-10-16T09:29:59Zpaperaa:paper_09291903_v11_n5_p333_FinocchiaroInstitucionalhttps://digital.bl.fcen.uba.ar/Universidad públicaNo correspondehttps://digital.bl.fcen.uba.ar/cgi-bin/oaiserver.cgiana@bl.fcen.uba.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:18962025-10-16 09:30:00.032Biblioteca Digital (UBA-FCEN) - Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturalesfalse
dc.title.none.fl_str_mv Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
title Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
spellingShingle Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
Finocchiaro, L.M.E.
Lipofection
Murine adenocarcinoma
Suicide gene therapy
ganciclovir
suicide substrate
thymidine kinase
animal cell
animal tissue
apoptosis
article
breast adenocarcinoma
bystander effect
cell growth
cell proliferation
cell specificity
cell type
controlled study
drug cytotoxicity
drug efficacy
gene expression regulation
gene transfer
Herpes simplex virus
in vitro study
in vivo study
intracellular transport
melanoma cell
mitochondrion
monolayer culture
mouse
multicellular spheroid
nonhuman
phenotype
population structure
priority journal
protein assembly
sensitivity analysis
statistical significance
transgene
virus resistance
Adenocarcinoma
Animals
Antiviral Agents
bcl-2-Associated X Protein
beta-Galactosidase
Bystander Effect
Cell Division
Cell Line, Tumor
Drug Resistance
Ganciclovir
Gene Therapy
Mammary Neoplasms, Animal
Melanoma
Mice
Mice, Inbred BALB C
Proto-Oncogene Proteins c-bcl-2
Simplexvirus
Spheroids, Cellular
Thymidine Kinase
Transfection
Viral Proteins
Animalia
Murinae
Simplexvirus
title_short Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
title_full Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
title_fullStr Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
title_full_unstemmed Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
title_sort Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
dc.creator.none.fl_str_mv Finocchiaro, L.M.E.
Bumaschny, V.F.
Karara, A.L.
Fiszman, G.L.
Casais, C.C.
Glikin, G.C.
author Finocchiaro, L.M.E.
author_facet Finocchiaro, L.M.E.
Bumaschny, V.F.
Karara, A.L.
Fiszman, G.L.
Casais, C.C.
Glikin, G.C.
author_role author
author2 Bumaschny, V.F.
Karara, A.L.
Fiszman, G.L.
Casais, C.C.
Glikin, G.C.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Lipofection
Murine adenocarcinoma
Suicide gene therapy
ganciclovir
suicide substrate
thymidine kinase
animal cell
animal tissue
apoptosis
article
breast adenocarcinoma
bystander effect
cell growth
cell proliferation
cell specificity
cell type
controlled study
drug cytotoxicity
drug efficacy
gene expression regulation
gene transfer
Herpes simplex virus
in vitro study
in vivo study
intracellular transport
melanoma cell
mitochondrion
monolayer culture
mouse
multicellular spheroid
nonhuman
phenotype
population structure
priority journal
protein assembly
sensitivity analysis
statistical significance
transgene
virus resistance
Adenocarcinoma
Animals
Antiviral Agents
bcl-2-Associated X Protein
beta-Galactosidase
Bystander Effect
Cell Division
Cell Line, Tumor
Drug Resistance
Ganciclovir
Gene Therapy
Mammary Neoplasms, Animal
Melanoma
Mice
Mice, Inbred BALB C
Proto-Oncogene Proteins c-bcl-2
Simplexvirus
Spheroids, Cellular
Thymidine Kinase
Transfection
Viral Proteins
Animalia
Murinae
Simplexvirus
topic Lipofection
Murine adenocarcinoma
Suicide gene therapy
ganciclovir
suicide substrate
thymidine kinase
animal cell
animal tissue
apoptosis
article
breast adenocarcinoma
bystander effect
cell growth
cell proliferation
cell specificity
cell type
controlled study
drug cytotoxicity
drug efficacy
gene expression regulation
gene transfer
Herpes simplex virus
in vitro study
in vivo study
intracellular transport
melanoma cell
mitochondrion
monolayer culture
mouse
multicellular spheroid
nonhuman
phenotype
population structure
priority journal
protein assembly
sensitivity analysis
statistical significance
transgene
virus resistance
Adenocarcinoma
Animals
Antiviral Agents
bcl-2-Associated X Protein
beta-Galactosidase
Bystander Effect
Cell Division
Cell Line, Tumor
Drug Resistance
Ganciclovir
Gene Therapy
Mammary Neoplasms, Animal
Melanoma
Mice
Mice, Inbred BALB C
Proto-Oncogene Proteins c-bcl-2
Simplexvirus
Spheroids, Cellular
Thymidine Kinase
Transfection
Viral Proteins
Animalia
Murinae
Simplexvirus
dc.description.none.fl_txt_mv We have developed multicellular spheroids (MCS) established from LM05e and LM3 spontaneous Balb/c-murine mammary adenocarcinoma and B16 C57-murine melanoma derived cell lines as an in vitro model to study the efficacy of the herpes simplex virus thymidine kinase/ganciclovir (HSVtk/GCV) suicide system. We demonstrated for the first time that HSVtk-expressing cells assembled as MCS manifested a GCV resistance phenotype compared to the same cells grown as sparse monolayers. HSVtk-expressing LM05e, LM3 and B16 spheroids were 16-, three- and nine-fold less sensitive to GCV than their respective monolayers, even though they could express transgenes 10-, eight- and five-fold more efficiently. Mixed populations of HSVtk- and their respective βgal-expressing cells displayed a cell-type specific bystander effect that was higher in monolayers than in MCS. However, HSVtk-expressing cells in two- or three-dimensional cultures were always significantly more sensitive to GCV than the βgal-expressing counterparts, supporting the feasibility of this suicide approach in vivo. We present evidence showing that HSVtk-expressing tumor cells, when transferred from monolayers to MCS, displayed: (i) lower GCV cytotoxic activity and bystander effect; (ii) higher and efficient expression of genes transferred as lipoplexes; (iii) lower cell proliferation rates; and (iv) changes in intracellular Bax/Bcl-xL rheostat of mitochondria-mediated apoptosis.
Fil:Finocchiaro, L.M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Casais, C.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Glikin, G.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
description We have developed multicellular spheroids (MCS) established from LM05e and LM3 spontaneous Balb/c-murine mammary adenocarcinoma and B16 C57-murine melanoma derived cell lines as an in vitro model to study the efficacy of the herpes simplex virus thymidine kinase/ganciclovir (HSVtk/GCV) suicide system. We demonstrated for the first time that HSVtk-expressing cells assembled as MCS manifested a GCV resistance phenotype compared to the same cells grown as sparse monolayers. HSVtk-expressing LM05e, LM3 and B16 spheroids were 16-, three- and nine-fold less sensitive to GCV than their respective monolayers, even though they could express transgenes 10-, eight- and five-fold more efficiently. Mixed populations of HSVtk- and their respective βgal-expressing cells displayed a cell-type specific bystander effect that was higher in monolayers than in MCS. However, HSVtk-expressing cells in two- or three-dimensional cultures were always significantly more sensitive to GCV than the βgal-expressing counterparts, supporting the feasibility of this suicide approach in vivo. We present evidence showing that HSVtk-expressing tumor cells, when transferred from monolayers to MCS, displayed: (i) lower GCV cytotoxic activity and bystander effect; (ii) higher and efficient expression of genes transferred as lipoplexes; (iii) lower cell proliferation rates; and (iv) changes in intracellular Bax/Bcl-xL rheostat of mitochondria-mediated apoptosis.
publishDate 2004
dc.date.none.fl_str_mv 2004
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/20.500.12110/paper_09291903_v11_n5_p333_Finocchiaro
url http://hdl.handle.net/20.500.12110/paper_09291903_v11_n5_p333_Finocchiaro
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by/2.5/ar
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/2.5/ar
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv Cancer Gene Ther. 2004;11(5):333-345
reponame:Biblioteca Digital (UBA-FCEN)
instname:Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
instacron:UBA-FCEN
reponame_str Biblioteca Digital (UBA-FCEN)
collection Biblioteca Digital (UBA-FCEN)
instname_str Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
instacron_str UBA-FCEN
institution UBA-FCEN
repository.name.fl_str_mv Biblioteca Digital (UBA-FCEN) - Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
repository.mail.fl_str_mv ana@bl.fcen.uba.ar
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