RADIATION-INDUCED DAMAGE TO DNA IN DRUG-RESISTANT AND RADIATION-RESISTANT SUBLINES OF A HUMAN BREAST-CANCER CELL-LINE

An Adriamycin-resistant subline of a human breast cancer cell line, MCF-7 ADR(R), has been shown to exhibit radioresistance associated with an increase in the size of the shoulder on the radiation survival curve. In the present study, damage to DNA of MCF-7 sublines WT and ADR(R) by 60Co γ radiation was measured by filter elution techniques. The initial amount of DNA damage, measured by both alkaline and neutral filter elution, was lower in ADR(R) cells, suggesting that these cells are resistant to radiation-induced single- and double-strand DNA breaks. In the case of double-strand breaks the difference between WT and ADR(R) cells was significant only at the lower radiation doses studied (up to 100 Gy). In cells depleted of glutathione (GSH) by L-buthionine sulfoximine (BSO) treatment, ADR(R) cells were sensitized to radiation-induced DNA damage, while WT cells were unaffected. The rate of repair of single- and double-strand DNA breaks following radiation was the same for both sublines, and repair of radiation damage was not affected by BSO treatment in either cell line. The relative resistance of ADR(R) cells to initial DNA damage by radiation is the only difference so far detected at the molecular level which reflects radiation survival, and it is possible that other factors are involved in the resistance of ADR(R) cells to killing by radiation. Sensitization of ADR(R) cells to radiation-induced DNA damage by GSH depletion, although not likely to involve inhibition of GSH- dependent detoxification enzymes per se (irradiation was done at 4°C), suggests that at the molecular level radioresponse in this subline is related to maintenance of GSH/GSSG redox equilibrium.