Objective      To construct a recombinant adenovirus vector of human trefoil factor family 3 (TFF3) and to determine its effect on the migration in human biliary epithelial cells.       Methods      The genomic fragment of target gene TFF3 was amplified with PCR with genomic DNA of liver cancer tissue as template. The obtained product was subcloned into adenovirus shuttle plasmid to construct vector pAdTrack-CMV-TFF3 after sequencing. Homologous recombination was performed by transfering the vector to E.coli BJ5183 containing t

he backbone pAd-Easy. The correct recombinant pAdEasy-TFF3 was selected and linearized with PacⅠ

then transfected into 293 cells for packaging and amplification. The titration of the recombinant adenovirus was determined. RT-PCR and Western blotting were used to determine the expression of TFF3 at mRNA and protein levels in human biliary epithelial cells after transfection. Finally

cell scratch test was performed to observe the effect on cell migration before and after transfection.       Results      The target fragment of TFF3 with 458 bp was successfully amplified

which was completely the same as the sequence published GenBank by DNA sequencing. The TFF3 homologous recombination adenovirus vector was successfully constructed with a titer of 1.3×10

9

CPU/mL. RT-PCR and Western blotting indicated significant increased expression of TFF3 at mRNA and protein levels in human biliary epithelial cells after transfection. Cell scratch test showed that cell migration was obviously activated and enhanced after transfection.       Conclusion      Recombinant adenovirus vector containing TFF3 gene is successfully constructed

which provides a satisfactory transduction vector to research the role of TFF3 in human biliary epithelium wound healing.