Anti-Angiogenesis by a Novel VEGF-Intrakine Strategy for Breast Cancer Therapy

摘要

Tumor angiogenesis plays an important role in breast cancer growth and metastasis. Vascular endothelial growth factor (VEGF) stimulates the proliferation of endothelial cells after binding to its receptor (VEGF- R), and is a key factor in tumor angiogenesis. The purpose of this project is to inactivate VEGF-R in vascular endothelial cells by using an intrakine/intrabody strategy to block the cell surface receptor expression intracellularly and thereby prevent endothelial cell proliferation. The scope of this research involves generating various expression vectors for the VEGF-intrakine/intrabody, determining their effects on the VEGF-R expression, developing an adeno- associated virus system for efficient transduction and evaluating the anti- angiogenesis activity in a mouse model. So far, immunoprecipitation, immunofluorescent and flow cytometry assays have been used to demonstrate that VEGF receptor-2 KDR is highly expressed on the surface of HUVEC. They also demonstrated the background KDR expression, which can be used as an important reference for future study of KDR expression after introducing VEGF- intrakine/ intrabody into the HUVEC cell line. VEGF intrakine/intrabody expression vectors have been generated by using RT-PCR and other gene construction methods. Now the intrakine/intrabody gene can be introduced into the cells and their anti- angiogenesis function will be studied in vitro and in vivo.