Characterization of a Chinese hamster ovary cell line with acquired resistance to the bisdioxopiperazine dexrazoxane (ICRF-187) catalytic inhibitor of topoisomerase II.
A Chinese hamster ovary (CHO) cell line highly resistant to the non-cleavable complex-forming topoisomerase II inhibitor dexrazoxane (ICRF-187, Zinecard) was selected. The resistant cell line (DZR) was 1500-fold resistant (IC50 = 2800 vs 1.8 microM) to continuous dexrazoxane exposure. DZR cells were also cross-resistant (8- to 500-fold) to other bisdioxopiperazines (ICRF-193, ICRF-154, and ICRF-186), and somewhat cross-resistant (4- to 14-fold) to anthracyclines (daunorubicin, doxorubicin, epirubicin, and idarubicin) and etoposide (8.5-fold), but not to the other non-cleavable complex-forming topoisomerase II inhibitors suramin and merbarone. The cytotoxicity of dexrazoxane to both cell lines was unchanged in the presence of the membrane-active agent verapamil. DZR cells were 9-fold resistant to dexrazoxane-mediated inhibition of topoisomerase II DNA decatenation activity compared with CHO cells (IC50 = 400 vs 45 microM), but were only 1.4-fold (IC50 = 110 vs 83 microM) resistant to etoposide. DZR cells contained one-half the level of topoisomerase II protein compared with parental CHO cells. However, the specific activity for decatenation using nuclear extract topoisomerase II was unchanged. Etoposide (100 microM)-induced topoisomerase II-DNA complexes in DZR cells and isolated nuclei were similarly one-half the level found in CHO cells and in isolated nuclei. However, the ability of 500 microM dexrazoxane to inhibit etoposide (100 microM)-induced topoisomerase II-DNA covalent complexes was reduced 4- to 6-fold in both DZR cells and nuclei compared with CHO cells and nuclei. In contrast, there was no differential ability of aclarubicin or merbarone to inhibit etoposide-induced topoisomerase II-DNA complexes in CHO compared with DZR cells and isolated nuclei. It was concluded that the DZR cell line acquired its resistance to dexrazoxane mainly through an alteration in the topoisomerase II target.
Hasinoff
B B
BB
Faculty of Pharmacy, University of Manitoba, Winnipeg, Canada. B_Hasinoff@UManitoba.ca
Kuschak
T I
TI
Creighton
A M
AM
Fattman
C L
CL
Allan
W P
WP
Thampatty
P
P
Yalowich
J C
JC
D015250:Aclarubicin / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells / Q000187:drug effects*; D006224:Cricetinae; D004250:DNA Topoisomerases, Type II / Q000235:genetics; D004351:Drug Resistance; D005029:Ethylenediamines / Q000633:toxicity; D005047:Etoposide / Q000037:antagonists & inhibitors / Q000494:pharmacology; D005998:Glycine / Q000031:analogs & derivatives / Q000633:toxicity; D009154:Mutation; D011929:Razoxane / Q000633:toxicity*; D013858:Thiobarbiturates / Q000494:pharmacology; D059005:Topoisomerase II Inhibitors; D014700:Verapamil / Q000494:pharmacology
D005029:Ethylenediamines; D013858:Thiobarbiturates; D059005:Topoisomerase II Inhibitors; D011929:Razoxane; D005047:Etoposide; D015250:Aclarubicin; C059866:ICRF 198; D014700:Verapamil; D004250:DNA Topoisomerases, Type II; D005998:Glycine; C045459:merbarone