title: Treatment with hydroxyurea and tyrphostin-1 significantly improves the transduction efficiency of recombinant adeno-associated viruses in human cancer cells.
abstract: To enhance the transduction efficiency (TE) of a recombinant adeno-associated virus 2 (rAAV2) in human cancer cells, we examined the combined effects of various chemicals known to influence the rAAV2 transduction process at distinct steps. Among the agents tested were trichostatin A, a histone deacetylase inhibitor, MG-132, a proteosome inhibitor, the genotoxic agents hydroxyurea, aphidicolin, etoposide and camptothecin, and tyrphostin-1, an epidermal growth factor receptor inhibitor. During or after chemical treatment, various human cancer cells were infected with rAAV2 expressing beta-galactosidase. Treatment with hydroxy-urea or etoposide plus tyrphostin-1 dramatically increased the TE in most cell lines. The combination of hydroxyurea plus tyrphostin-1 increased TE to 37.7+/-7.9%, 32.8+/-2.0% and 31.8+/-2.1% in SK-Hep1, HeLa, and HCT116 cells, respectively. In addition, following rAAV2 infection and treatment with hydroxyurea plus tyrphostin-1, long-term transgene expression was observed for up to 6 months, with no damage to the transduced cells. These results indicate that rAAV2 transgene expression can be significantly enhanced by a combination of chemical agents with distinct activity and prolonged gene expression can occur following rAAV2 gene transfer into human cancer cells.
authors.affiliation: Department of Microbiology, University of Ulsan College of Medicine, Songpa P.O. Box 145, Seoul 138-736, Korea.
authors.forename: Sung Jin
authors.initials: SJ
authors.lastname: Kim
authors.forename: Young Ran
authors.initials: YR
authors.lastname: Nam
authors.forename: Ohkyu
authors.initials: O
authors.lastname: Shin
authors.forename: Jene
authors.initials: J
authors.lastname: Choi
authors.forename: Boyoung
authors.initials: B
authors.lastname: Lee
authors.forename: Jin Woo
authors.initials: JW
authors.lastname: Chang
authors.forename: Yunhee Kim
authors.initials: YK
authors.lastname: Kwon
authors.forename: Keerang
authors.initials: K
authors.lastname: Park
authors.forename: Heuiran
authors.initials: H
authors.lastname: Lee
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology*; D015151:Immunoblotting; D007763:Lac Operon / Q000235:genetics; D009369:Neoplasms / Q000235:genetics / Q000473:pathology; D013997:Time Factors; D014162:Transfection; D020032:Tyrphostins / Q000494:pharmacology*; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D004274:DNA, Recombinant; D020032:Tyrphostins; D001616:beta-Galactosidase; D006918:Hydroxyurea