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Abstract:

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.

Registro:

Documento: Artículo
Título:Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
Autor:Finocchiaro, L.M.E.; Bumaschny, V.F.; Karara, A.L.; Fiszman, G.L.; Casais, C.C.; Glikin, G.C.
Filiación:Unidad de Transferencia Genetica, Inst. de Oncologia Angel H. Roffo, Universidad de Buenos Aires, Buenos Aires, Argentina
Unidad de Transferencia Genetica, Inst. de Oncologia Dr AH Roffo - UBA, Av. San Martín 5481, 1407 Buenos Aires, Argentina
INGEBI-CONICET/FCEN-UBA, Argentina
Palabras clave: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
Año:2004
Volumen:11
Número:5
Página de inicio:333
Página de fin:345
DOI: http://dx.doi.org/10.1038/sj.cgt.7700682
Título revista:Cancer Gene Therapy
Título revista abreviado:Cancer Gene Ther.
ISSN:09291903
CODEN:CGTHE
CAS:ganciclovir, 82410-32-0; thymidine kinase, 9002-06-6, 9086-73-1; Antiviral Agents; Bax protein, mouse; Ganciclovir, 82410-32-0; Proto-Oncogene Proteins c-bcl-2; Thymidine Kinase, 2.7.1.21; Viral Proteins; bcl-2-Associated X Protein; beta-Galactosidase, 3.2.1.23
PDF:https://bibliotecadigital.exactas.uba.ar/download/paper/paper_09291903_v11_n5_p333_Finocchiaro.pdf
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09291903_v11_n5_p333_Finocchiaro

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Citas:

---------- APA ----------
Finocchiaro, L.M.E., Bumaschny, V.F., Karara, A.L., Fiszman, G.L., Casais, C.C. & Glikin, G.C. (2004) . Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids. Cancer Gene Therapy, 11(5), 333-345.
http://dx.doi.org/10.1038/sj.cgt.7700682
---------- CHICAGO ----------
Finocchiaro, L.M.E., Bumaschny, V.F., Karara, A.L., Fiszman, G.L., Casais, C.C., Glikin, G.C. "Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids" . Cancer Gene Therapy 11, no. 5 (2004) : 333-345.
http://dx.doi.org/10.1038/sj.cgt.7700682
---------- MLA ----------
Finocchiaro, L.M.E., Bumaschny, V.F., Karara, A.L., Fiszman, G.L., Casais, C.C., Glikin, G.C. "Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids" . Cancer Gene Therapy, vol. 11, no. 5, 2004, pp. 333-345.
http://dx.doi.org/10.1038/sj.cgt.7700682
---------- VANCOUVER ----------
Finocchiaro, L.M.E., Bumaschny, V.F., Karara, A.L., Fiszman, G.L., Casais, C.C., Glikin, G.C. Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids. Cancer Gene Ther. 2004;11(5):333-345.
http://dx.doi.org/10.1038/sj.cgt.7700682