[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.


CONCLUSIONS
The results indicated that the truncated VEGF-C cDNA was sufficient for expression, secretion and activation when it was transfected into Tca8113 cell.
Hua
Cheng-ge
CG
Department of Oral and Maxillofacial Surgery, West China Stomatological Hospital, Sichuan University, Chengdu 610041, China.
Pan
Jian
J
Wen
Yu-ming
YM
Li
Min
M
Wang
Gang
G
Zhang
Yi-zheng
YZ
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
D018076:DNA, Complementary; D042582:Vascular Endothelial Growth Factor C