Effect of adenovirus mediated human intestinal trefoil factor gene on intestinal epithelial cells

Abstract

Objective To construct a recombinant adenovirus vector of human intestinal trefoil factor(hITF)and observe its effect on human intestinal epithelial cells(Caco-2). Methods A recombinant adenovirus vector containing hITF gene was constructed and infected Caco- 2 cells. The expression of hITF mRNA and protein were detected by reverse transcriotion-polymerase chain reaction(RT-PCR), Western blotting and immunofluorescence. Then, wounds were established in confluent monolayers of Caco-2 cells infected with recombinant adenovirus vector, and the width of the cells migration was measured in a blind manner. Results The recombinant adenovirus vector Ad-hITF was constructed and the infection efficiency was about 100%. RT- PCR and Western blotting assay demonstrated that hITF was transcribed and expressed successfully. hITF was proved to distribute in cytoplasm by immunofluorescence assay. In addition, Ad-hITF enhanced migration activity of HT-29 cells obviously. Conclusion Recombinant adenovirus vector containing hITF gene was successfully constructed and its effect was confirmed in promoting cells migration. This research lay foundations for the further application of hITF gene therapy. Key words: Intestinal trefoil factor; Adenovirus; Intestinal epithelial cells; Migration