title: Effect of anEDA-A1gene mutant on the proliferation and cell cycle distribution of cultured human umbilical vein endothelial cells.
abstract: Ectodysplasin (EDA) gene mutation is associated with hypohidrotic ectodermal dysplasia (HED). The aim of this study was to investigate the effect of ectodysplasin, transcript variant 1 (EDA-A1) on the proliferation and cell cycle of ECV304 human umbilical vein endothelial cells (HUVECs). Recombinant eukaryotic expression vectors containing mutant (M) and wild-type (W)EDA-A1coding sequences, pcDNA3.1 (-)-EDA-A1-M and pcDNA3.1 (-)-EDA-A1-W, respectively, were transfected into ECV304 cells. The EDA-A1 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR), and the protein was detected by western blotting. The EDA-A1 gene and protein were detected in ECV304 cells transfected with pcDNA3.1 (-)-EDA-A1-M and pcDNA3.1 (-)-EDA-A1-W, but not in ECV304 cells transfected with empty plasmid or cells that had not undergone transfection. Compared with the control group, the EDA-A1 gene mutant significantly decreased the proliferation of ECV304 cells and its inhibition rate was 45.70% (P<0.01), whereas the wild-type EDA-A1 gene did not cause such growth inhibition (P>0.05). A significant increase of the fraction of cells in the G0/G1phase of the cell cycle was observed in the ECV304 cells of the mutant group compared with wild type group, with an increase in the S phase population and a concomitant reduction in the G2/M phase population (P<0.05). These results indicate that compared with the wild-type gene, transfection with a mutant EDA-A1 gene inhibited the proliferation and cell cycle of cultured HUVECs.
authors.affiliation: Department of Stomatology, Central Hospital of Guangyuan, Guangyuan, Sichuan 628000, P.R. China.
authors.forename: K E
authors.initials: KE
authors.lastname: Lei
authors.affiliation: Department of Stomatology, Central Hospital of Guangyuan, Guangyuan, Sichuan 628000, P.R. China.
authors.forename: Lunchang
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Respiratory Medicine, Central Hospital of Guangyuan, Guangyuan, Sichuan 628000, P.R. China.
authors.forename: Bing
authors.initials: B
authors.lastname: Ma
authors.affiliation: Department of Respiratory Medicine, Central Hospital of Guangyuan, Guangyuan, Sichuan 628000, P.R. China.
authors.forename: Ping
authors.initials: P
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
authors.forename: Longjiang
authors.initials: L
authors.lastname: Li
authors.affiliation: Institute of Cell Biology, School of Life Science, Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
authors.forename: Tuanjie
authors.initials: T
authors.lastname: Che
authors.affiliation: Department of Prosthodontics, School of Stomatology, Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
authors.forename: Xiangyi
authors.initials: X
authors.lastname: He
keywords: cell cycle; ectodysplasin; human umbilical vein endothelial cell; mutant; proliferation; transcript variant 1 gene