Synopsis
The effect of 4′-demethyl-epipodophyllotoxin-β-d-thenylidene glucoside (VM-26), a semi-synthetic derivative of podophyllotoxin, on the cell cycle was studied with chick embryo fibroblasts cultivatedin vitro. DNA, RNA and protein content, as well as NADH-diaphorase activity were determined by quantitative microdensitometry and cytofluorometry. The incorporation of [3H]thymidine and [3H]leucine into DNA and proteins were analysed by autoradiography. These metabolic data correlated with morphological observation showed that VM-26 blocks the cell cycle at different moments of its kinetics depending on both the dose and the time exposure. NADH-diaphorase activity is the first to be affected, then biochemical changes (involving the metabolism of RNA and proteins) and morphological alterations (especially of mitochondria) follow. This suggests that VM-26 may act primarily upon the mechanism of respiration of the cell.
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References
Appleton, T. C. (1972). Stripping film autoradiography. In:Autoradiography for Biologists (ed. P. B. Gahan), pp. 33–49. London, New York: Academic Press.
Benthley, R. (1861). New American remedies. 1: Popophyllum pellatum.Phar. J. Trans. 3, 456.
Bullough, W. S. (1952). Energy relations of mitotic activity.Biol. Rev. 27, 133–68.
Butcher, R. G. (1972). Precise cytochemical measurement of neotetrazolium formazan by scanning and integrating microdensitometry.Histochemie 32, 171–90.
Butcher, R. G. &Altman, F. P. (1973). Studies on the reduction of tetrazolium salts. II- The measurement of the half reduced and fully reduced formazans of neotetrazolium chloride in tissue sections.Histochemie 37, 351–63.
Caspersson, T. &Lomakka, G. (1970). Recent progress in quantitative cytochemistry: instrumentation and results. In:Introduction to Quantitative Cytochemistry, Vol. II (eds. G. L. Wied and G. F. Bahr), pp. 27–56. London, New York: Academic Press.
Crissman, H. A. &Steinkamp, J. A. (1973). Rapid simultaneous measurement of DNA, protein and cell volume in single cells from large mammalian cell populations.J. Cell Biol. 59, 766–71.
Deitch, A. D. (1966). Cytophotometry of proteins. In:Introduction to Quantitative Cytochemistry (ed. G. L. Wied), pp. 451–68. New York, London: Academic Press.
Epel, D. (1963). The effects of carbon monoxide inhibition on ATP level and the rate of mitosis in the sea-urchin egg.J. Cell Biol. 17, 315–9.
Gautschi, J. R., Schindler, R. &Hurni, C. (1971). Studies on the division cycle of mammalian cells. V. Modifications of time parameters by different steady-state culture conditions.J. Cell Biol. 51, 653–63.
Goldsmith, M. A. &Carter, S. K. (1973). 4′-Demethyl-epipodophyllotoxin-β-d-thenylidne glucoside (VM-26). A brief review.Europe. J. Cancer. 9, 477–82.
Gosalvez, M., Blanco, M., Hunter, J., Miko, M. &Chance, B. (1974). Effects of anticancer agents on the respiration of isolated mitochondria and tumor cells.Europ. J. Cancer. 10, 567–74.
Gosalvez, M., Perez-Garcia, J. &Lopez, M. (1972). Inhibition of NADH-linked respiration with the anti-cancer agent 4′-demethyl-epipodophyllotoxin thenylidene glucoside (VM-26).Europ. J. Cancer. 8, 471–3.
Gorzos, V. (1977). Histochemical study of nucleic acids and proteins by cytophotometry and histoautoradiography in fibroblasts cultivated in hyperoxia.Acta histochem. 58, 141–55.
Gotzos, V., Spreca, A. &Conti, G. (1971). Action de O-(β-hydroxyaethyl)-Rutosidea (HR) sur les fibroblastes de l'embryon de poulet cultivésin vitro.Arch. Anat. microsc. Morph. exp. 60, 133–45.
Mak, S. (1965). Mammalian cell cycle analysis using microspectrophotometry combined with autoradiography.Exp. Cell Res. 39, 286.
Mallet, M. (1975).Abrége de cytologie. p. 37. Paris: Masson.
Mazia, D. (1961). Mitosis and the physiology of cell division. D. Identification of specific preparations for division. In:The Cell. Vol. II (eds. J. Brachet and A. E. Mirsky), pp. 108–147. New York, London: Academic Press.
Ruch, F. (1970). Principles and some applications of cytofluorometry. In:Introduction of Quantitative Cytochemistry (eds. G. L. Wied), pp. 295–323. New York, London: Academic Press.
Sandritter, W., Kiefer, G. &Rick, W. (1966). Gallocyanin chrome alum. In:Introduction to Quantitative Cytochemistry, (ed. G. L. Wied), pp. 295–323. New York, London: Academic Press.
Stahelin, H. (1970). 4′-demethyl-epipodophyllotoxin-thenylidene glucoside (VM-26). A podophyllum compound with a new mechanism of action.Europ. J. Cancer 6, 303–11.
Swann, M. M. (1953). The mechanism of cell division. A study with carbon monoxide on the sea-urchin egg.Q. Jl microsc. Sci. 94, 369–79.
Taylor, E. (1973). Macromolecular assembly inhibitors and their action on the cell cycle. In:Drugs and the Cell Cycle (eds. A. M. Zimmerman, G. A. Padilla and I. L. Cameron), pp. 11–24. New York, London: Academic Press.
Tobey, R. A., Petersen, D. F. &Anderson, E. C. (1971). Biochemistry of G2 and mitosis. In:The Cell Cycle and Cancer (ed. R. Baserga), pp. 309–353. New York: Marcel Dekker.
Vendrely, C. (1971). Cytophotometry and histochemistry of the cell cycle. In:The Cell Cycle and Cancer (ed. R. Baserga), pp. 227–68. New York: Marcel Dekker.
Zetterberg, A. &Killander, D. (1965). Quantitative cytochemical studies on interphase growth. II. Derivation of synthesis curves from the distribution of DNA, RNA and mass values of individual mouse fibroblastsin vitro.Expl. Cell Res. 39, 22–32.
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Dedicated to Professor G. Conti on the occasion of his 60th birthday.
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Gotzos, V., Cappelli-Gotzos, B. & Despond, J.M. A quantitative microdensitometric and autoradiographic study of the effect of 4′-demethyl-epipodophyllotoxin-β-D-thenylidene glucoside (VM-26) on the cell cycle of cultured fibroblasts. Histochem J 11, 691–707 (1979). https://doi.org/10.1007/BF01004733
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DOI: https://doi.org/10.1007/BF01004733