Development and properties of an etoposide-resistant human leukaemic CCRF-CEM cell line.
Etoposide (VP-16) and several other unrelated anti-tumour agents appear to act by inhibiting the enzyme DNA topoisomerase II. We report here the development and characterization of an etoposide-resistant human leukaemic CCRF-CEM cell line, CEM/VP-1. The cell line was 15-fold more resistant to etoposide than the parental CEM cells and exhibited cross-resistance to other topoisomerase II inhibitors including teniposide, m-AMSA, and doxorubicin. CEM/VP-1 cells exhibited only a low level cross-resistance to the Vinca alkaloids, vinblastine and vincristine, known inhibitors of mitotic spindle formation. As a first step in defining the mechanism of resistance to etoposide, we compared the levels of topoisomerase II activity and its drug sensitivity in nuclear extracts from the resistant and sensitive CEM cells. As determined by a kinetoplast DNA decatenation assay, the level of DNA topoisomerase II activity in CEM/VP-1 nuclear extracts was approximately 2-fold lower than that in CEM cells, and the activity appeared to be resistant to inhibition by etoposide. Furthermore, the DNA topoisomerase II activity in CEM/VP-1 nuclear extracts did not promote the etoposide-dependent cleavage of pBR322 DNA observed with extract from sensitive cells. These results suggest that etoposide resistance in the CEM/VP-1 cell line may at least in part be due to an altered topoisomerase II, or associated factor, resulting in a reduced ability to induce DNA cleavage in the presence of drug.
Patel
S
S
Department of Cellular and Molecular Sciences, St George's Hospital Medical School, University of London, UK.
Austin
C A
CA
Fisher
L M
LM
D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000201:enzymology; D003470:Culture Media; D004250:DNA Topoisomerases, Type II / Q000032:analysis; D004270:DNA, Circular / Q000032:analysis / Q000187:drug effects; D018105:DNA, Kinetoplast; D004273:DNA, Neoplasm / Q000032:analysis / Q000187:drug effects; D004351:Drug Resistance; D005047:Etoposide / Q000037:antagonists & inhibitors* / Q000633:toxicity; D006801:Humans; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy* / Q000201:enzymology; D059005:Topoisomerase II Inhibitors; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000201:enzymology
D003470:Culture Media; D004270:DNA, Circular; D018105:DNA, Kinetoplast; D004273:DNA, Neoplasm; D059005:Topoisomerase II Inhibitors; D005047:Etoposide; D004250:DNA Topoisomerases, Type II