Quantitative and qualitative aspects of topoisomerase I and II alpha and beta in untreated and platinum/cyclophosphamide treated malignant ovarian tumors.
Quantitative and qualitative aspects of topoisomerase (Topo) I and II were studied in 17 malignant ovarian tumors [eight untreated and nine after platinum/cyclophosphamide (Pt/Cy) chemotherapy]. Median Topo II catalytic activity was lower (P < 0.05) in tumors after Pt/Cy chemotherapy in comparison to untreated tumors, while no differences were found for Topo I catalytic activity in tumors before and after chemotherapy, as was also found in a previous study (Van der Zee et al. Cancer Res., 51: 5915-5920, 1991). Teniposide (VM-26)-induced cleavable complex formation correlated (r = 0.60; P < 0.05) with Topo II activity, while Topo II decatenation activity was equally but incompletely inhibited by VM-26 in all tumors. No differences were found in Topo II cleavage site patterns in plasmid BR322 DNA for all tumors using an indirect end-labeling procedure. Cleavable complex formation of Topo I by camptothecin (Cpt) did not correlate with Topo I catalytic activity, while Topo I catalytic activity could equally and completely be inhibited by Cpt. By Western blotting, Topo II alpha protein expression was detected in four of eight untreated tumors and three of nine tumors after Pt/Cy chemotherapy, whereas in all tumors a M(r) 150,000 degradation product of Topo II beta was detected. Topo I protein was detected in all tumors at varying levels, but the protein levels did not correlate with Topo I catalytic activity or cleavable complex formation by Cpt. Our study shows that Topo I and II, isolated from human malignant tumors, can be stimulated by Cpt and VM-26, respectively, to induce DNA cleavage, which suggests that topoisomerases are real targets for chemotherapy in patients with ovarian cancer. From in vitro data from the literature it appears that the cleavable complex assay reflects both quantitative and qualitative changes as well as changes in the phosphorylation state of Topo I and II. In combination with the feasibility of the cleavable complex assay for Topo I and II in human malignant tumors, which was found in the present study, it appears that at present the determination of cleavable complex formation by tumors seems to be the most promising parameter of Topo I or II expression in human tumors to be related to response to Topo I- or II-targeted chemotherapy.
van der Zee
A G
AG
Department of Gynecology, University Hospital, Groningen, The Netherlands.
de Jong
S
S
Keith
W N
WN
Hollema
H
H
Boonstra
H
H
de Vries
E G
EG
D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use*; D015153:Blotting, Western; D002453:Cell Cycle / Q000502:physiology; D003520:Cyclophosphamide / Q000008:administration & dosage; D004249:DNA Damage; D004264:DNA Topoisomerases, Type I / Q000032:analysis / Q000378:metabolism; D004250:DNA Topoisomerases, Type II / Q000032:analysis / Q000378:metabolism; D004273:DNA, Neoplasm / Q000032:analysis / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009944:Organoplatinum Compounds / Q000008:administration & dosage; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000201:enzymology* / Q000473:pathology; D013713:Teniposide / Q000494:pharmacology; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors
D004273:DNA, Neoplasm; D007527:Isoenzymes; D009944:Organoplatinum Compounds; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors; D003520:Cyclophosphamide; D013713:Teniposide; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II