Resistance of p53 knockout cells to doxorubicin is related to reduced formation of DNA strand breaks rather than impaired apoptotic signaling.
The anthracycline doxorubicin (adriamycin) is an important chemotherapeutic agent used in the treatment of solid epithelial and mesenchymal tumors as well as leukemias. A variety of mechanisms has been proposed to be involved in doxorubicin-induced cytotoxicity such as DNA intercalation, oxidative stress, DNA strand breakage by inhibition of topoisomerase II, activation of death receptors, and altered p53 expression. Concerning doxorubicin resistance and p53 status data reported are contradictory. Here, we show that mouse fibroblasts deficient in p53 (p53(-/-)) are more resistant to doxorubicin than p53 wild-type (p53 wt) cells. This is in contrast to other genotoxic agents (UV-light, alkylating drugs) for which p53(-/-) fibroblasts proved to be more sensitive. Resistance of p53(-/-) cells to doxorubicin is related to reduced induction of apoptosis. This is not likely to be due to altered apoptotic signaling since the expression of Bax and Bcl-2 was unchanged and the induction of Fas/CD95/APO-1 receptor and caspase-8 was the same in p53(-/-) and p53 wt cells on treatment with doxorubicin. However, we observed a clearly lower level of doxorubicin-induced DNA strand breaks in p53(-/-) cells compared to the wt. P170 glycoprotein was equally expressed and the accumulation and elimination of the drug occurred with identical kinetics in both cell types. p53 deficient cells were cross-resistant to another topoisomerase II inhibitor etoposide, which also provoked increased DNA strand breakage in p53 wt cells. Based on the data we conclude that the p53 status significantly impacts the generation of DNA strand breaks because of drug-induced topoisomerase inhibition rather than death receptor signaling. Since human tumors are frequently mutated in p53 the findings bear clinical implications.
Dunkern
Torsten R
TR
Division of Applied Toxicology, Institute of Toxicology, University of Mainz, Obere Zahlbacher Strasse 67, D-55131 Mainz, Germany.
Wedemeyer
Inga
I
Baumgärtner
Manuela
M
Fritz
Gerhard
G
Kaina
Bernd
B
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002460:Cell Line; D004249:DNA Damage / Q000502:physiology*; D004317:Doxorubicin / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D066298:In Vitro Techniques; D051379:Mice; D018345:Mice, Knockout; D015398:Signal Transduction / Q000502:physiology; D059005:Topoisomerase II Inhibitors; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism*
D059005:Topoisomerase II Inhibitors; D016159:Tumor Suppressor Protein p53; D004317:Doxorubicin