Radiosensitive human tumour cell lines show misrepair of DNA termini

Physical measures of the rejoining of radiation-induced breaks in DNA strands are limited in terms of sensitivity and the fact that they do not assess the fidelity with which the rejoining occurs. In this report, transfection of cleaved plasmid has been used as a probe for repair in three radiosensitive tumour cell lines and shown them to have low repair fidelity compared with resistant cells. Errors in the repair of linear plasmid were found by Southern analysis, in keeping with the measured repair fidelity. Radiosensitive tumour cells showed few errors in the uptake and integration of circular plasmid, in contrast to ataxia-telangiectasia (A-T) cells. In the neuroblastoma HX142, the repair of blunt-ended linear plasmid was associated with deletions of > 1 kb; staggered-ended linear plasmid was repaired with small insertions and circular plasmid integration was intact in > 60% of the copies. The neuroblastoma SKN.SH, processed staggered-ended plasmid by insertions of a variety of sizes, but processed circular plasmid largely error-free. In contrast, A-T cells (AT5BIVA) had the same spectrum of errors irrespective of the form of plasmid transfected. Cell fusion between HX142 and AT5BIVA showed complementation to a resistant phenotype, suggesting that misrepair in the tumour cell did not result from somatic mutation in the ATM gene. In conclusion, radiosensitive tumours show evidence of misrepair of DNA termini, with a mechanism which is functionally and genetically distinct from that in A-T cells.