Treatment with hydroxyurea and tyrphostin-1 significantly improves the transduction efficiency of recombinant adeno-associated viruses in human cancer cells.
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.
Kim
Sung Jin
SJ
Department of Microbiology, University of Ulsan College of Medicine, Songpa P.O. Box 145, Seoul 138-736, Korea.
Nam
Young Ran
YR
Shin
Ohkyu
O
Choi
Jene
J
Lee
Boyoung
B
Chang
Jin Woo
JW
Kwon
Yunhee Kim
YK
Park
Keerang
K
Lee
Heuiran
H
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
D004274:DNA, Recombinant; D020032:Tyrphostins; D001616:beta-Galactosidase; D006918:Hydroxyurea