APPARENT ALTERATIONS IN THE EARLY-STAGE OF EXCISION-REPAIR OF UV-INDUCED DNA DAMAGES IN A HELA MUTANT-CELL LINE THAT IS RESISTANT TO GENOTOXIC STRESSES

We have previously reported a cisplatin-resistant HeLa cell line featuring a cross-resistance to genotoxic stresses including ultraviolet (UV) radiation and an enhancement of plasmid reactivation. In this study, excision repair of UV-DNA adducts in this resistant cell line was investigated. Using alkaline elution analysis, this resistant cell line showed a 2-fold enhancement in damage incision-associated DNA strand breaks. Using a gel mobility shift assay, the resistant cells exhibited a 3-fold increase in nuclear proteins which specifically recognize UV-damaged DNA. However, the rate of repair synthesis in the resistant cells appeared to be the same as in their parental counterparts. Thus, recognition and incision activities, the early stage of excision repair, are altered in the resistant cells. The results suggest that phenotypic cross-resistance of this cell line to UV is probably due to an improved excision repair of UV-DNA adducts which is defective in xeroderma pigmentosum group A cells. The results are consistent with the conclusion that the early stage, including recognition and incision, of excision repair is critical in determining cellular sensitivity or resistance to DNA damage.