Boulaiz Houria, Prados José, Marchal Juan Antonio, García Angel Miguel, Alvarez Luis, Melguizo Consolación, Carrillo Esmeralda, Ramos Juan Luis, Aránega Antonia
Basic Cardiovascular Research Section, Department of Morphological Sciences, School of Medicine, University of Granada, E-18012 Granada, Spain.
Cancer Sci. 2003 Jun;94(6):564-8. doi: 10.1111/j.1349-7006.2003.tb01483.x.
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.
在大肠杆菌DNA中发现的gef基因编码一种小蛋白(50个氨基酸),该蛋白与细胞杀伤功能有关。我们使用MS - 36黑色素瘤细胞系作为实验模型,来检验gef基因作为一种癌症治疗新策略的有效性。我们使用pMAMneo载体转染MS - 36细胞,并用地塞米松诱导gef基因表达。与MS - 36亲本细胞相比,这使MS - 36TG的增殖率降低了多达85%。细胞生长的减少伴随着细胞周期和形态的显著改变。G1期逐渐消失,细胞积聚在S期。然而,用膜联蛋白V - FITC和7 - 氨基放线菌素D进行的研究未能证明诱导了细胞凋亡。形态学变化包括细胞大小增加、丝状伪足数量增多,尤其是细胞膜上出现了亲本细胞中未见的孔状改变。我们的结果表明,gef基因是一种独立于前药给药的系统,能显著降低MS - 36细胞的增殖。因此,该基因可能被认为是癌症基因治疗的一个新候选基因。