Construction of pcDNA3-Endo eukaryon expression plasmid and angiogenesis inhibition in vitro / 中国组织工程研究

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 groupsrecombinant 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.