Transfection of MS-36 melanoma cells with gef gene inhibits proliferation and induces modulation of the cell cycle.
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.
Boulaiz
Houria
H
Basic Cardiovascular Research Section, Department of Morphological Sciences, School of Medicine, University of Granada, E-18012 Granada, Spain. hboulaiz@ugr.es
Prados
José
J
Marchal
Juan Antonio
JA
García
Angel Miguel
AM
Alvarez
Luis
L
Melguizo
Consolación
C
Carrillo
Esmeralda
E
Ramos
Juan Luis
JL
Aránega
Antonia
A
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
D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; C071524:HokC protein, E coli