Construction of Hepatocyte Growth Factor Gene Recombinant Adenovirus Vector and its Expression in Vascular Smooth Muscle Cells

Aim To construct an adenovirus expression vector which can express hepatocyte growth factor (HGF) in vascular smooth muscle cells (SMC). Methods The plasmid containing HGF fragment was cleaved by restriction enzyme digestion, and the resultant fragment was inserted directionally into adenoviral shuttle plasmid. The linearized recombinant adeno viral shuttle plasmid and adenovirus expression vector were cotransformed into Escherichia coli BJ5183 cells for homologous recombination . The resultant recombinant plasmid, pAd-HGF, then was transfected into HEK293 cells with liposome for packaging. The recombinant adenoviral shuttle plasmid and pAd-HGF were identified by enzyme digestion and sequencing. The package of pAd-HGF in HEK293 cells was tracked by fluorescent microscope, and was observed by electronic microscope. The expression of packaged pAd-HGF in abdominal aortic SMCs of rat was identified by RT-PCR and Western blotting. Results HGF fragment was inserted correctly into the adenoviral shuttle plasmid and adenovirus expression vector. High-liter packaged adenovirus vector was produced and expressed in abdominal aortic SMCs of rat. Conclusions A recombinant adenovirus expression vector of HGF was constructed successfully. This study suggested that HGF may be a potential target for the gene therapy of vascular diseases and established a foundation for further study.