title: [Construction of RBM5 vector, establishment of stably transfected A549 cell line and preliminary research on the function of RBM5 gene].
abstract: OBJECTIVE
  To establish a stable A549 cell line transfected by RNA binding motif 5 (RBM5) expression vector, and to investigate the effect of RBM5 gene on proliferation of A549 cell line and the expression of DEAH box polypeptide 15 (DHX15).
  
  
  METHODS
  The eukaryotic expression vector pcDNA3.1 (+)/RBM5 was constructed by a twostep PCR technique. Then, the recombinant plasmid pcDNA3.1 (+)/RBM5 was verified by DNA sequencing and transfected into the lung adenocarcinoma cell A549. The positive cells with overexpression of RBM5 gene were identified by Western blotting. Flow cytometry was used to analyze the cell cycles of the positive A549 cells [pcDNA3.1 (+)/RBM5-A549] and the negative controls [pcDNA3 .1 (+)- A549]. Finally, RT-PCR was used to detect the expression of DHX15, a splicing-related factor, in the positively transfected A549 cells and the negative controls.
  
  
  RESULTS
  A pcDNA3.1 (+)/RBM5 eukaryotic expression vector has been constructed successfully, and the A549 cell line that stably transfected with RBM5 gene has been established. Compared with negative control cells, the percentage of G1 phase cells in the positive cells was increased, while the percentage of S phase was decreased (both P<0.01), and the expression of DHX15 is upregulated (P<0.01).
  
  
  CONCLUSIONS
  RBM5 gene can inhibit the cell cycle and upregulate the expression of DHX15 in A549 cells.
authors.affiliation: Department of Geriatrics, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Xiao
authors.forename: Nian
authors.initials: N
authors.lastname: Li
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Xing
authors.forename: Bixiu
authors.initials: B
authors.lastname: He
mesh_terms: D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004268:DNA-Binding Proteins / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D020365:RNA Helicases / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics*; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000235:genetics*
chemical_list: D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C473448:RBM5 protein, human; D016601:RNA-Binding Proteins; D025521:Tumor Suppressor Proteins; C539469:DHX15 protein, human; D020365:RNA Helicases