Treatment with the anti-tumor drugs, cis-platin and mafosfamide, does not affect the structure of prekinetochores in a human breast cancer cell line. An immunofluorescence study using human anti-centromere autoantibodies.
The goal of the present article was to determine whether a nuclear parameter, centromere structure of interphase cells, could serve as an indicator to assess cellular damage caused by anti-tumor drugs. These were cis-platin and mafosfamide, which are widely used for the management of solid tumors. To visualize the centromeres, we probed treated and untreated cells of a human breast cancer cell line, MX-1, with a human anti-centromere serum. The serum was obtained from a scleroderma patient and detects antigens associated with prekinetochores of the decondensed chromosomes. The DNA was simultaneously displayed by a specific fluorescent dye. The cells were grown on coverslips, incubated for 1 h in a drug-containing medium, transferred into a drug-free medium and observed 24 h later. Since the efficiency of many anti-tumor drugs increases with the temperature, two different temperatures, 37 and 42 degrees C, were used. The analysis revealed that the treatment did not visibly alter the labeling pattern. We conclude that chromosome structure remains largely intact and is not suitable for the cytological evaluation of the efficiency of anti-tumor drugs. This is in contrast with, for example, the microtubular cytoskeleton and mitochondria, which were extensively damaged under the conditions applied here (compare Wolf et al. 1995). Independent of the drug and the temperature selected, the nuclear lumen of mononucleated and multinucleated cells contained small fluorescent spots. Double dots corresponding to the sister centromeres in the G2 phase of the cell cycle were rare. In addition to the voluminous nuclei, some cells possessed micronuclei in the lateral cytoplasm and these were regularly labeled by the autoantibodies. A small subset of the mononucleated MX-1 cells had unusually large nuclei. It is reasonable to assume that they are polyploid. The fluorescent spots marking the prekinetochores were very large in these cells. This may indicate that the chromosomes remain associated after replication.
Wolf
K W
KW
Institut für Anthropologie, Universität Mainz, Deutschland.
Mentzel
M
M
Mendoza
A S
AS
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000473:pathology; D002503:Centromere / Q000187:drug effects / Q000648:ultrastructure; D002945:Cisplatin / Q000494:pharmacology*; D003520:Cyclophosphamide / Q000031:analogs & derivatives* / Q000494:pharmacology; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D018386:Kinetochores / Q000187:drug effects*; D048629:Micronuclei, Chromosome-Defective / Q000187:drug effects; D008854:Microscopy, Electron; D013696:Temperature; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; C048341:mafosfamide; D003520:Cyclophosphamide; D002945:Cisplatin