[The combination of VEGF-C expressing vector and the detection of expression in Tca8113 cell].

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. CONCLUSION: The results indicated that the truncated VEGF-C cDNA was sufficient for expression, secretion and activation when it was transfected into Tca8113 cell.