title: [Construction of γ-synuclein eukaryotic expression vector and its effect on invasion and metastasis of colon cancer cell line SW1116 in vitro].
abstract: OBJECTIVE
  To construct γ-synuclein gene eukaryotic expression vector, and to study its effect on the invasion of colon cancer cell line SW1116 and the adhesion between SW1116 and human umbilical vein endothelial cells(HUVECs) in vitro.
  
  
  METHODS
  Total RNA was extracted from colon cancer cell line HT29 and the cDNA of γ-synuclein was amplified using RT-PCR. The digested fragment of cDNA coding sequence was linked to the eukaryotic expression vector pEGFP-C1 containing the GFP gene. After identification by sequence analysis, the recombinant plasmid was transfected into colon cancer cell line SW1116 via lipofectamine. The stable cell line was selected with G-418. The invasion in vitro was tested by Transwell invasion chamber assay. HUVECs were previously seeded onto 96-well plates before SW1116 cells seeded, and fluorescence intensity of GFP was detected to represent the amount of adhesion cells by ELISA.
  
  
  RESULTS
  Human γ-synuclein eukaryotic expression vector was successfully constructed, which was stably expressed in SW1116 cells and could translate the GFP-γ-synuclein protein in vitro. γ-synuclein facilitated SW1116 cell passing through matrigel and filter membrane(198.4±20.7 vs. 98.8±13.2, P<0.05) and elevated the adherence of SW1116 cells to HUVECs(3.08±0.36 vs. 1.22±0.21, P<0.05).
  
  
  CONCLUSIONS
  Expression of γ-synuclein can strengthen colon cancer cell SW1116 potentiality of invasion and metastasis in vitro.
authors: Department of Abdominal Surgery, Fujian Provincial Tumor Hospital, Fujian Medical University, Fujian Key Laboratory of Tumor Biological Treatment, Fuzhou 350014, China. surghuangfeng@yahoo.com.cn.
authors: Qing
authors: Q
authors: Ye
authors: Feng
authors: F
authors: Huang
authors: Qiuhong
authors: Q
authors: Zheng
authors: Xiaoying
authors: X
authors: Wang
authors: Yangmei
authors: Y
authors: Xu
authors: Fusheng
authors: F
authors: Gong
authors: Lijie
authors: L
authors: Huang
mesh_terms: D002448:Cell Adhesion; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000473:pathology*; D005822:Genetic Vectors / Q000096:biosynthesis*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D051845:gamma-Synuclein / Q000235:genetics*
chemical_list: D051845:gamma-Synuclein