title: Efficient Adenovirus Gene Transfer Methods in Human Colonic Caco-2 Epithelial Cells Using Capric Acid.
abstract: Adenovirus (Ad) vectors are widely used in gene therapy and in vitro/in vivo gene transfer. However, Ad-mediated gene transfer in epithelial cells shows low efficiency, because Ad fiber cannot bind to the primary receptor, the coxsackievirus and adenovirus receptor (CAR), present in tight junctions. Caco-2 monolayer cells cultured on Transwell-chamber plates for approximately 2 weeks are widely used for drug membrane permeation studies, but Ad-mediated gene transfer is difficult in Caco-2 monolayer cells. First, we examined the efficiency of gene transfer into Caco-2 monolayer cells. Luciferase production in cultured Caco-2 cells transduced with Ad vectors was 20-fold lower on day 12 than on day 1. In contrast, the expression of CAR protein in Caco-2 cells gradually increased along with the duration of culture. For efficient gene transfer into Caco-2 monolayer cells, the binding ability of Ad vectors with CAR was found to be important. Capric acid (C10), a medium-chain fatty acid is a tight-junction modulator used as a pharmaceutical agent. We found that a novel gene transfer method using transduction with Ad vectors in the presence of C10 led more efficiently to LacZ expression in Caco-2 monolayer cells than Ad vectors alone. The results of the present study indicate that C10 could be very useful for Ad-mediated gene transfer in human colonic Caco-2 epithelial cells.
authors.affiliation: Department of Pharmaceutics and Biopharmaceutics, Showa Pharmaceutical University.
authors.forename: Naoya
authors.initials: N
authors.lastname: Koizumi
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Yamagishi
authors.forename: Takamasa
authors.initials: T
authors.lastname: Hirai
authors.forename: Makiko
authors.initials: M
authors.lastname: Fujii
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Mizuguchi
authors.forename: Yoshiteru
authors.initials: Y
authors.lastname: Watanabe
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000257:Adenoviridae Infections / Q000378:metabolism; D018938:Caco-2 Cells; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein / Q000378:metabolism*; D003652:Decanoic Acids / Q000494:pharmacology*; D004847:Epithelial Cells / Q000378:metabolism*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D019108:Tight Junctions; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D003652:Decanoic Acids; C031071:decanoic acid; D008156:Luciferases