Temperature dependence of adriamycin-induced DNA damage in L1210 cells.
We report alkaline elution experiments that reveal the temperature dependence of DNA lesions, both single-strand breaks and DNA-protein cross-links, in L1210 cells exposed to Adriamycin. DNA damage, which at 37 degrees C is equivalent to several hundred rads of ionizing radiation exposure, diminishes as the temperature of drug exposure is lowered. At all temperatures below about 15 degrees C no DNA damage is detectable in L1210 cells exposed to Adriamycin, even at relatively high doses. The low temperature inactivity is not due to a redistribution of intracellular drug since at both 37 and 0 degrees C there is a high concentration of Adriamycin in both nuclear and cytoplasmic locations. The temperature profile for DNA damage parallels the profile for cytotoxicity, i.e., at low temperature, the drug is completely inactive as a cytotoxic agent (P. Lane, P. Vichi, D. L. Bain, and T. R. Tritton, Cancer Res., 47:4038-4042, 1987). Thus, DNA breaks and cell kill appear to be correlated with one another. However, when we examined DNA lesions in nuclei isolated from L1210 cells we found that the low temperature inability to sustain Adriamycin-induced single-strand breaks or DNA-protein cross-links was absent. In nuclei, then, the drug can provoke DNA damage at low temperature, while in whole cells it cannot. Topoisomerase II, an enzyme implicated in catalyzing DNA lesions in cells exposed to intercalating agents, retains its catalytic activity both to unknot P4 DNA at 0 degrees C, and to be induced by drug to alter the release of pBR322 supercoils, so a low temperature inactivation of this enzyme cannot explain the results. We propose that intact L1210 cells have a regulatory factor which controls DNA damage, possibly through topoisomerase II, but which is lost when nuclei are isolated.
Vichi
P
P
Department of Pharmacology, University of Vermont College of Medicine, Burlington.
Robison
S
S
Tritton
T R
TR
D000818:Animals; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004249:DNA Damage; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D004317:Doxorubicin / Q000378:metabolism / Q000633:toxicity*; D066298:In Vitro Techniques; D007939:Leukemia L1210; D051379:Mice; D010957:Plasmids; D013696:Temperature; D059005:Topoisomerase II Inhibitors
D059005:Topoisomerase II Inhibitors; D004317:Doxorubicin; D004250:DNA Topoisomerases, Type II