Construction of pcDNA3-Endo eukaryon expression plasmid and angiogenesis inhibition in vitro
Shao Jia-jia
Yu Yin
Jiang Tao
Abstract：BACKGROUND:The eradication therapy of glioma is the major problem, and anti-angiogenesis therapy is a potential treatment of glioma. <br> 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. <br> 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. <br> 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.
Keywords：endostatinsendothelial cellsneovascularizationphysiologicmesenchymal stem cells
Publication Date：2014-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 4636-4641 )