Construction of pcDNA3-Endo eukaryon expression plasmid and angiogenesis inhibition in vitro
10.3969/j.issn.2095-4344.2014.29.008
- VernacularTitle:体外构建内皮抑素基因真核表达载体抑制血管生成
- Author:
Jiajia SHAO
;
Yin YU
;
Tao JIANG
- Publication Type:Journal Article
- Keywords:
endostatins;
endothelial cells;
neovascularization,physiologic;
mesenchymal stem cells
- From:
Chinese Journal of Tissue Engineering Research
2014;(29):4636-4641
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:The eradication therapy of glioma is the major problem, and anti-angiogenesis therapy is a potential treatment of glioma.
OBJECTIVE:To confirm the inhibiting effect of endostatin on angiogenesis in vitro, and to lay the foundation in inhibiting the growth of tumor by endostatin in the future.
METHODS:Endostatin mRNA was extracted from the liver of Wistar rats by Trizol and endostatin cDNA was synthesized by RT-PCR. Endostatin cDNA and pcDNA3 were connected and pcDNA3-Endo recombined plasmid was constructed successful y. The recombinant pcDNA3-Endo was transfected into bone marrow mesenchymal stem cells by Lipofectamine. The expression of endostatin was identified by RT-PCR and western blot analysis. Endostatin proteinum activity was detected by ECV-304 cellproliferation inhibition experiment using MTT assay. The in vitro experiments were divided into four groups:recombinant plasmid group, vector plasmid group, liposome control group and blank control group.
RESULTS AND CONCLUSION:PcDNA3-Endo eukaryon expression plasmid was constructed successful y. Endostatin gene can be transcribed and expressed effectively in vitro by pcDNA3-Endo plasmid. The growth of ECV-304 cellwas inhibited obviously by pcDNA3-Endo. The growth of vascular endothelial cells can be inhibited obviously by endostatin gene.
