Enhancement of DNA-transfection frequency by X-rays.
This study was conducted to evaluate the frequency of DNA transfection into human cells following X-ray irradiation. We transfected plasmid DNA (pSV2neo) into human cells, HeLa and PA-1, by either calcium phosphate precipitation or the lipofection method immediately after irradiating the cells with various doses of X-rays. The transfection frequency was evaluated by counting the number of G418-resistant colonies. When circular plasmid DNA was used, irradiation up to a dose of 2 Gy dose-dependently increased the transfection frequency, which reached a maximum of 5 to 10-fold that of the control unirradiated cells. When linear plasmid DNA was used, the transfection frequency was 2 times higher than that of circular DNA. All five of the clones that were randomly chosen expressed the transfected neo gene. In addition, the pSV2neo gene was randomly integrated into the genomic DNA of each clone. These findings indicate that X-ray treatment can facilitate foreign DNA transfer into human cells and that radiation-induced DNA breaks may promote the insertion of foreign DNA into host DNA. The enhancement of DNA transfection with X-rays may be instrumental in practicing gene therapy.
Iwamoto
R
R
Department of Radiology, Okayama University Medical School, Japan.
Fushimi
K
K
Hiraki
Y
Y
Namba
M
M
D002460:Cell Line; D004247:DNA / Q000235:genetics* / Q000528:radiation effects; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids / Q000235:genetics*; D011829:Radiation Dosage; D014162:Transfection / Q000235:genetics* / Q000528:radiation effects
D004247:DNA