Inhibitory effects of Ad-VEGI151 on venous endothelial cell proliferation / 第二军医大学学报

ABSTRACT

Objective:To study the inhibitory effects of adenovirus mediated vascular endothelial cell growth inhibitor 151(VEGI_(151)) on venous endothelial cell proliferation. Methods: VEGI_(151) gene was cloned into adenovirus vector pCA13 and the product was packaged and amplified with 293A cells.The expressed protein was identified by Western blot. X-gal staining was used to determine the transfection efficiency of the recombinant adenovirus system. The inhibitory effect of Ad-VEGI_(151) on ECV304 cell proliferarion was observed and assessed by violet crystal assay. The protein expression of VEGI_(151) gene in ECV304 cells was detected by immunohistochemistry. Results: Liposome-mediated pCA13-VEGI_(151) and pJM17 cotransfection of 293A cells (through homologous recombination of intracellular plasmid) is a practical and feasible method to obtain high-titre recombinant adenovirus. The constructed Ad-VEGI_(151), with high transfection efficiency, significantly inhibited ECV304 cells proliferation and expressed functional protein in target cells. Conclusion: Ad-VEGI_(151) can express bioactive protein in target cells and can inhibit the proliferation of venous endothelial cells, and this may pave a new way for gene therapy of tumors and neovascular diseases.