Recombinant hFGF-10 adenovirus promotes proliferation of kerotinocytes

AIM:To construct the recombinant adenoviral vector containing human fibroblast growth factor 10(hFGF-10) gene,and to study the effect of the recombinant adenovirus on the proliferation of kerotinocytes.METHODS: HFGF-10 gene was amplified by PCR and ligated with shuttle vector pAdTrack-CMV to get the recombinant plasmid pAdTrack-CMV-hFGF-10,which was linearized with PmeI and transferred into Escherichia coli BJ5183 containing the adenoviral bone plasmid pAdEasy-1 for homologous recombination to obtain the recombinant adenoviral plasmid pAdEasy-hFGF-10.The recombinant adenoviral plasmid was then transfected into HEK-293 cell line to package and amplify the recombinant adenovirus.The expression of hFGF-10 in HaCat cells infected with the recombinant adenovirus was detected by Western blotting.The influence of the recombinant adenovirus on the proliferation of kerotinocytes was checked by MTT.RESULTS: The recombinant adenovirus containing hFGF-10 gene was successfully constructed,which effectively infected HaCat cells.The result of Western blotting showed that a protein in culture media of the infected HaCat cells reacted with hFGF-10 antibody.The recombinant adenovirus stimulated the proliferation of kerotinocytes.CONCLUSION: HaCat cells infected with the recombinant adenovirus expresses and secrets hFGF-10 protein,which promotes the proliferation of HaCat cells.