title: Transfection of MS-36 melanoma cells with gef gene inhibits proliferation and induces modulation of the cell cycle.
abstract: The gef gene, found in Escherichia coli DNA, encodes a small (50 amino acids) protein which is related to cell-killing functions. We used the MS-36 melanoma cell line as an experimental model to examine the usefulness of the gef gene as a new strategy for cancer therapy. We transfected MS-36 cells using the pMAMneo vector, and induced gef gene expression with dexamethasone. This decreased the proliferation rate of MS-36TG by as much as 85% in comparison with MS-36 parental cells. The decrease in cell growth was accompanied with significant modifications of the cell cycle and morphology. The G1-phase gradually disappeared, with accumulation in the S-phase. However, studies with annexin V-FITC and 7-aminoactinomycin D failed to demonstrate induction of apoptosis. Morphological changes were an increase in cell size and the number of filopodia, and especially the appearance of pore-like alterations in the cell membrane which were not seen in parental cells. Our results demonstrate that the gef gene, a system independent of the administration of a prodrug, significantly reduces the proliferation of MS-36 cells. This gene may therefore be considered a new candidate for cancer gene therapy.
authors.affiliation: Basic Cardiovascular Research Section, Department of Morphological Sciences, School of Medicine, University of Granada, E-18012 Granada, Spain. hboulaiz@ugr.es
authors.forename: Houria
authors.initials: H
authors.lastname: Boulaiz
authors.forename: José
authors.initials: J
authors.lastname: Prados
authors.forename: Juan Antonio
authors.initials: JA
authors.lastname: Marchal
authors.forename: Angel Miguel
authors.initials: AM
authors.lastname: García
authors.forename: Luis
authors.initials: L
authors.lastname: Alvarez
authors.forename: Consolación
authors.initials: C
authors.lastname: Melguizo
authors.forename: Esmeralda
authors.initials: E
authors.lastname: Carrillo
authors.forename: Juan Luis
authors.initials: JL
authors.lastname: Ramos
authors.forename: Antonia
authors.initials: A
authors.lastname: Aránega
mesh_terms: D017209:Apoptosis; D001427:Bacterial Toxins / Q000235:genetics* / Q000378:metabolism; D002455:Cell Division; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D016193:G1 Phase; D015316:Genetic Therapy; D006801:Humans; D008545:Melanoma / Q000378:metabolism / Q000473:pathology*; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016196:S Phase; D012878:Skin Neoplasms / Q000378:metabolism / Q000473:pathology*; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; C071524:HokC protein, E coli