Study on construction of a plasmid vector carrying human hepatocyte growth factor gene and activity of its expression product.
- Author:
Xiao-Qin HA
1
;
Xin-Guo WANG
;
Zu-Ze WU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cells, Cultured; DNA, Complementary; genetics; Genetic Vectors; Hepatocyte Growth Factor; genetics; metabolism; Humans; Plasmids; Rats; Rats, Wistar; Transfection
- From: Chinese Journal of Applied Physiology 2002;18(3):278-282
- CountryChina
- Language:Chinese
-
Abstract:
AIMTo construct a plasmid carrying hepatocyte growth factor gene and investigate its effects in vitro.
METHODSA complementary DNA (cDNA) clone for human HGF was isolated from human placental cDNA, then subcloned into pUDK vector, which was constructed by ourselves, to form the pUDKH plasmid. The transfection efficiency and the expression level of HGF and VEGF were evaluated by transfecting pUDK or pUDKH into primary rat skeletal muscle cells. The biological effects of HGF-expressing product at different doses on endothelial cells were investigated in vitro, and assessed by MTT.
RESULTSThe primary rat skeletal muscle cells could be transfected efficiently with pUDKH (0.057%), and secreted HGF(16 -18 ng/4 x 10(5) cells) and VEGF proteins. The expressing product could significantly stimulate proliferation of human umbilical vein endothelial cells, in a dose-dependent manner (P < 0.05).
CONCLUSIONpUDKH has the potential application in vivo to treat ischemic diseases.
