title: [The combination of VEGF-C expressing vector and the detection of expression in Tca8113 cell].
abstract: OBJECTIVE
  To investigate the functional expression of a truncated VEGF-C gene in mammalian cells.
  
  
  METHODS
  A truncated VEGF-C cDNA (with 5'-terminal cleaved between residues 207/208) was used to construct a pcDNA3.1(+)-VEGF-C recombined plasmid, which was cloned into a lingual SCC cell line (Tca8113) by lipid-mediated transfection. The expression of VEGF-C was detected by Western blotting and immunohistochemical staining.
  
  
  RESULTS
  The VEGF-C fragment was successfully ligated into the pcDNA3.1(+) between EcoR I and Xho I multiclone sites. By Western blotting, the relative molecular mass 53 x 10(3) and 29 x 10(3) forms of VEGF-C were detected in cell lysates of VEGF-C transfected cells (VcTca), while the relative molecular mass 29 x 10(3) form of VEGF-C presented the only band in culture media. According to immunohistochemistry, the VcTca cells showed intensive cytoplasm positive reaction to VEGF-C, whereas faint reaction was found in non-transfected cells and pcDNA3.1(+) mock transfected cells.
  
  
  CONCLUSIONS
  The results indicated that the truncated VEGF-C cDNA was sufficient for expression, secretion and activation when it was transfected into Tca8113 cell.
authors.affiliation: Department of Oral and Maxillofacial Surgery, West China Stomatological Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Cheng-ge
authors.initials: CG
authors.lastname: Hua
authors.forename: Jian
authors.initials: J
authors.lastname: Pan
authors.forename: Yu-ming
authors.initials: YM
authors.lastname: Wen
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.forename: Yi-zheng
authors.initials: YZ
authors.lastname: Zhang
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D011387:Prokaryotic Cells / Q000378:metabolism; D014062:Tongue Neoplasms / Q000378:metabolism / Q000473:pathology; D014162:Transfection; D042582:Vascular Endothelial Growth Factor C / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D042582:Vascular Endothelial Growth Factor C