Ultrasound facilitates transduction of naked plasmid DNA into colon carcinoma cells in vitro and in vivo.
One approach to improve the efficacy of in vivo gene therapy, with the aim at enhancing expression of the transgene, involves utilization of mechanical forces to facilitate transduction of DNA into cells. In this study, we evaluated the feasibility of mechanical insonation in gene transfers with naked DNA plasmid loading both in vitro and in vivo. We used an ultrasound probe, which can focus the ultrasonic beam in the exit zone of the probe. The reporter pcDNA3-lacZ plasmid, containing Escherichia coli lacZ or the beta-galactosidase gene (beta-gal), and the neomycin 3'-phosphotransferase gene (neo), was used for evaluation of transfer efficiency. Expression of beta-gal in MC38 murine colon carcinoma cells was measured after insonation of 20 W/cm2 with continuous 1.0-MHz wave exposure. In a transient assay, significant numbers of cells were transduced with the beta-galactosidase gene. After cells were treated with geneticin, we also observed a difference in colonogenicity between noninsonated and insonated groups. When MC38 cells were implanted in syngeneic mice and plasmid was injected, the insonation that followed facilitated beta-galactosidase expression. These results indicate that insonation represents a potential approach for gene therapy when combined with naked DNA plasmid injection.
Manome
Y
Y
Department of Microbiology, Jikei University School of Medicine, Tokyo, Japan.
Nakamura
M
M
Ohno
T
T
Furuhata
H
H
D000230:Adenocarcinoma; D000818:Animals; D003110:Colonic Neoplasms; D018014:Gene Transfer Techniques / Q000295:instrumentation; D005822:Genetic Vectors; D019868:Kanamycin Kinase / Q000235:genetics*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids; D014407:Tumor Cells, Cultured; D014465:Ultrasonics; D001616:beta-Galactosidase / Q000235:genetics*
D019868:Kanamycin Kinase; D001616:beta-Galactosidase