Construction of pcDNA3-Endo eukaryon expression plasmid and angiogenesis inhibitionin vitro

    loading  Checking for direct PDF access through Ovid

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 successfully. 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 cell proliferation 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 successfully. Endostatin gene can be transcribed and expressed effectively in vitro by pcDNA3-Endo plasmid. The growth of ECV-304 cell was inhibited obviously by pcDNA3-Endo. The growth of vascular endothelial cells can be inhibited obviously by endostatin gene.

Subject headings:

endostatins; endothelial cells; neovascularization, physiologic; mesenchymal stem cells

Related Topics

    loading  Loading Related Articles