title: [Establishment of an N-2a cell line stably expressing mouse galanin and the effect of over-expressed galanin on the proliferation and apoptosis of N-2a cell].
abstract: OBJECTIVE
  To construct an N-2a cell line stably expressing PcDNA 3.1-platelet derived growth factor-galanin (GAL) and explore the effect of over-expressed GAL on proliferation and apoptosis of N-2a cell in vitro.
  
  
  METHODS
  The vector containing the target gene was transfected into N-2a cells by liposome, and cell clones stably over-expressing GAL was obtained via G418 screening. GAL mRNA and protein levels were determined by reverse transcriotion-polymerase chain reaction (RT-PCR) and Western blot. The proliferation of N-2a cells was detected by MTT.The cell cycle and apoptosis were detected by flow cytometry.
  
  
  RESULTS
  RT-PCT and Western blot indicated that GAL genes were highly expressed in the transfected N-2a cells (i.e.GAL-N-2a). As shown by MTT, the proliferation of the N-2a cells transfected with PcDNA 3.1-PDGF-GAL was significantly slower than the control group(P<0.05). Compared with the non-transfected cells in the control group, the N-2a cells with endogenously overexpressed GAL were arrested at the G0/G1 phases, and the over-expressed GAL protein significantly induced the N-2a cell apoptosis in a concentration-dependent fashion.
  
  
  CONCLUSIONS
  Eukaryotic expression vector PcDNA 3.1-PDGF-GAL can encode the expression of GAL in N-2a cells. Aslo, it can inhibit cell proliferation and promote the cell apoptosis.
authors.affiliation: Laboratory Animal Center of Shanxi Medical University, Taiyuan 030001, China.
authors.forename: Man
authors.initials: M
authors.lastname: Rong
authors.forename: Rui-hu
authors.initials: RH
authors.lastname: Zhang
authors.forename: Tian-fu
authors.initials: TF
authors.lastname: Liu
mesh_terms: D000818:Animals; D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019004:Galanin / Q000096:biosynthesis*; D051379:Mice; D012333:RNA, Messenger / Q000096:biosynthesis
chemical_list: D012333:RNA, Messenger; D019004:Galanin