Summary
C6 glioma cells (CCL 107) were cultured for three days and then treated withcis-dichlorodiamineplatinum (cis-DDP) at doses of 0.2–10 µg/ml medium. Changes in DNA synthesis and DNA content, as well as morphology of cells and chromatin distribution, were examined from the first post-treatment day onwards. The number of cells labelled with [3H]thymidine, detected autoradiographically, decreased after treatment with 0.2–10 µg/ml by approximately one half on post-treatment day 1 and diminished further by the third day after treatment. The labelled cells were entirely absent only after treatment with 10µg/ml, 7 days post-treatment. Mitoses decreased from 1.4–0.6% by post-treatment day 1 and completely disappeared by day 3 (1 µg/ml). Feulgen cytophotometry and propidium iodide cytofluorimetry revealed accumulation of cells in the S-phase, especially the latter part (0.5 and 1.0 µg/ml, post-treatment day 1) and subsequently also in G2 phase (post-treatment day 3). Incomplete cyto- and karyokinesis in some cycling cells was indicated by an increased number of binucleate cells and nuclei of higher ploidy classes. Labelled cells with intermediate DNA values were, on average, labelled less intensively, as was revealed by simultaneous measurements of DNA content and [3H]thymidine incorporation. Some cells displayed reduction in grain density over heterochromatin clumps. This would be in agreement with the late S-phase block of DNA replication. After post-treatment day 3 the density of cells in cultures was substantially lower. This was due to slowed transversing through the cell cycle and cell death occurring after post-treatment day 1 with higher doses or after day 2 with lower doses (up to 1 µg/ml). The size of the nuclei of surviving cells enlarged initially (post-treatment day 1) and later (day 7) giant cells with long, branched fibres similar to those of reactive astrocytes occurred. Texture analysis of Feulgen-stained nuclei revealed that the chromatin of cells treated withcis-DDP became less evenly distributed. This might be due either to the direct influence ofcis-DDP on the DNA molecule, or mediated by changes in cytoskeleton and cAMP levels described earlier.
Similar content being viewed by others
References
AGGARWAL, S. K. (1974) Inhibition of cytokinesis in mammalian cells bycis-dichloro-diamine-platinum (II).Cytobiologie 8, 395–402.
BERGERAT, J. A., BARLOGIE, B., COEHDE, W., JOHNSTON, D. A. & DREWINKO, B. (1979)In vitro cytokinetics response of human colon cancer cells tocis-dichlorodiamineplatinum (II).Cancer Res. 39, 4356–63.
BILLINGS, P. C., ORF, J. W., PALMER, D. K., TALMAGE, D. A., PAN, C. G. & BLUMENFELD, M. (1979) Anomalous electrophoretic mobility ofDrosophila phosphorylated H1 histone: is it related to the compaction of satellite DNA into heterochromatin?Nucl. Acid Res. 6, 2151–64.
D'AMATO, F. (1977) The cell cycle. InNuclear Cytology in Relation to Development p. 30. Cambridge: Cambridge University Press.
DROBNIK, J. (1983) Antitumour activity of platinum complexes.Cancer Chemoter. Pharmacol. 10, 145–9.
FLEISCHMANNOVÁ, V., SCHMEER, A., DROBNIK, J., LODIN, Z. & MAREŠ, V. (1982) The influence of drugs with a cytostatic action on the growth and differentiation of glioma cells.Physiol. Bohemosl. 31, 441.
FOLBERGROVÁ, J., LISÁ, V., DROBNIK, & MAREŠ, V. (1987) Cyclic AMP levels of C6 glioma cells treated withcis-dichlorodiamineplatinum (cis-DDP).Neoplasma 34, 3–16.
FREITAS, I., GIORDANO, P. A. & BOTTIROLI, G. (1981) Improvement in microscope photometry by voltage to frequency conversion: analogue measurement and digital processing.J. Microsc. 124, 211–18.
GIORDANO, P. A., MAZZINI, G., RICCARDI, A., MONTECUCCO, G., UCCI, G. & DANOVA, M. (1985) Propidium iodide staining of cytoautoradiographic preparations for the simultaneous determination of DNA content and grain count.Histochem. J. 17, 1259–70.
HOUSSIER, C., DEPAUW-GILLET, M. C., HACHA, R. & FREDERICQ, E. (1983) Alteration in the nucleosome and chromatin structures upon interaction with platinum coordination complexes.Biochim. Biophys. Acta 739, 317–25.
KUENZLE, C. C. (1985) Enzymology of DNA replication and repair in the brain.Brain Res. Rev. 10, 231–45.
LAZO, J. S., PRASAD, K. N. & RUDDON, R. W. (1976) Synthesis and phosphorylation of chromatin associated proteins in cAMP-induced ‘differentiated’ neuroblastoma cells in culture.Exp. Cell Res. 100, 41–6.
MAREŠ, V., FLEISCHMANNOVÁ, V., LODIN, Z. & UERERBERG, H. (1981) Cyclic AMP and growth regulation in rat glioma cells in tissue culture.Exp. Neurol. 71, 154–60.
MARKS, P. A., SHEFFREY, R., RAMSEY, R. & RIFKIND, F. A. (1986) Inducer mediated modulation of gene expression during murine erythroleukemia (MELC) differentiation, IUCC Proceedings, Karger Akademia Kiado, 1986, p. 1847.
MOONEN, G., CAM, Y., SENSENBRENNER, M. & MANDEL, P. (1975) Variability of the effects of serum free medium, db-cAMP or theophyline on the morphology of cultured newborn rat astroblasts.Cell Tissue Res. 163, 365–72.
OREDSSON, S. M., DEEN, D. F. & MARTON, L. J. (1982) Decreased cytotoxicity ofcis-diaminechloroplatinum (II) byα-difluoromethyl-ornithine depletion of polyamines in 9L rat brain.Cancer Res. 42, 1296–9.
REPTA, A. J. & LONG, D. F. (1980) Reactions ofcis-platinum with human plasma fractions. InCisplatin: Current Status and New Developments (edited by PRESTAYKO A. W., CROOKE, S. T. and CARTER, S. K.) p. 285. New York: Academic Press.
ROBERTS, J. J. & PERA, M. P. (1983) Action of platinum antitumour drugs. InMolecular Aspects of Anti-Cancer Drug Action. (edited by NEIDLE, S. and WARING, M. J.) p. 183–232. London: Macmillan.
SODHI, A. & AGGARWAL, S. K. (1974) Effects ofcis-dichloroamineplatinum (II) in the regression of sarcoma 190: a fine structural study.J. Nat. Cancer Inst. 53, 85–9.
SZUMIEL, I. & NIAS', A. H. W. (1976) Action of a platinum complexcis-dichlorobis(cyclopentylamine)-platinum (II) on Chinese hamster ovary cellsin vitro.Chem. Biol. Interact. 14, 217–32.
TACHACAROF, E., NATCHEV, T. C. H., MITRANY, L. & BOEV, K. (1965) Analyse discriminative de la non-homogeneite optique du noyau cellulaire.Mikriskopie 20, 331–5.
van EYS, J. (1985) Medical and oncological management of pediatric brain tumours.Prog. exp. Tumor. Res. 29, 235–48.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mareš, V., Giordano, P.A., Mazzini, G. et al. Influence ofcis-dichlorodiamineplatinum on glioma cell morphology and cell cycle kinetics in tissue culture. Histochem J 19, 187–194 (1987). https://doi.org/10.1007/BF01680628
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01680628