The stable expression of human tissue-type plasminogen activator gene mediated by lipofectamine in human vein endothelial cell line cells.
- Author:
Xiaobin LIU
1
;
Kailun ZHANG
;
Xionggang JIANG
;
Jianing WANG
;
Yongzhang HUANG
Author Information
1. Department of Cardiovascular Surgery, Guizhou Provincial Hospital, Guiyang 550002, China. liuxiaobin930@yahoo.com.cn
- Publication Type:Journal Article
- MeSH:
Cell Line;
Human Umbilical Vein Endothelial Cells;
metabolism;
Humans;
Lipids;
pharmacology;
RNA, Messenger;
biosynthesis;
genetics;
Recombinant Proteins;
biosynthesis;
genetics;
Tissue Engineering;
Tissue Plasminogen Activator;
biosynthesis;
genetics;
Transfection
- From:
Journal of Biomedical Engineering
2007;24(6):1330-1333
- CountryChina
- Language:Chinese
-
Abstract:
We have established a human umbilical vein endothelial cell (HUVEC) line monoclonal cells with the stable expression of human tissue-type plasminogen activator (t-PA) gene to provide a basis for further study on the vascular tissue engineering. Recombinant plasmid pcDNA3. 1-Myc-His B (-)/t-PA was constructed by insertion of t-PAcDNA originated from PBS/t-PA into eukaryotic expression vector pcDNA3. 1-Myc-His B(-) and transfected into hUVEC line cells mediated by lipofectamine. The positive clones were obtained by the screen of G418. The transcription and expression of t-PA gene were investigated by RT-PCR and Western blotting respectively. The t-PA activity was measured by chromogenic substrate assay. The positive clone cells which transcripted the mRNA of t-PA gene was obtained by RT-PCR. Immunoreactive human t-PA of the medium was significantly increased in the group of transfected gene when compared with that in the controlled and transfected plasmid without t-PA gene group. The biological activity of the protein of the t-PA in the media was increased significantly in the positive clone cells with t-PA gene transfected. The HUVEC line monoclonal cells with the stable expression of t-PA gene was established successfully.
