Summary
In contrast to normal liver cells, AS-30D hepatoma cells are insensitive to phalloidin. The lack of the typical phalloidin response in the latter cells is not due to a deficiency of contractile proteins. Actin prepared from hepatoma cells is able to form filamentous structures and is stabilized in a manner similar to muscle actin.
Isolated liver cells were exposed to a medium containing phalloidin and removed after 20 min by centrifugation. The supernatant was inculated again with fresh cells. The procedure was repeated four times. The phalloidin response decreased to about 19% of the control because of the uptake of phalloidin during each incubation. When the same procedure was carried out with AS-30D hepatoma cells, and aliquots of the supernatants were tested with hepatocytes no marked decrease of the phalloidin response was seen. This indicates that hepatoma cells do not consume the toxin as do normal liver cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Agostini, B., Hofmann, W.: Morphology of phalloidin intoxication. In: Membrane alterations as basis of liver injury (H. Popper, L. Bianchi, W. Reutter, eds.), pp. 277–292. Lancaster: MTP Press Ltd. 1977
Agostini, B., Govindan, V. M., Hofmann, W.: Morphological changes induced by phalloidin in the rat liver. In: Pathogenesis and mechanisms of liver cell necrosis (E. Keppler, ed.), pp. 175–192. Lancaster: MTP Press Ltd. 1975
Agostini, B., Wieland, Th., Ivankovic, S., Hofmann, W.: Phalloidin tolerance in rats with liver carcinoma induced by diethylnitrosamine. Naturwissenschaften 63, 438 (1976)
Agostini, B., Ivankovic, S., Hofmann, W.: Different response to phalloidin in rats with various hepatomas. Naturwissenschaften 64, 650 (1977)
Berry, M. N., Friend, D. S.: High-yield preparation of isolated rat liver parenchymal cells. J. Cell Biol. 43, 506–529 (1969)
Bischoff, E., Tran-Thi, T. A., Decker, K. F. A.: Nucleotide pyrophosphatase of rat liver. Eur. J. Biochem. 51, 353–361 (1975)
Clarke, M., Spudich, J. A.: Biochemical and structural studies of actomyosin-like proteins of non-muscle cells. J. Mol. Biol. 86, 209–222 (1974)
Dancker, P., Löw, I., Hasselbach, W., Wieland, Th.: Interaction of actin with phalloidin: polymerisation and stabilization of Factin. Biochim. Biophys. Acta 400, 407–414 (1975)
Faulstich, H., Wieland, Th., Schimassek, H., Walli, A. K., Ehler, N.: Mechanism of phalloidin intoxication. II. Binding studies. In: Membrane alterations as basis of liver injury (H. Popper, L. Bianchi, W. Reutter, eds.), pp. 301–310 Lancaster: MTP Press Ltd. 1977
Frimmer, M., Homann, J., Petzinger, E., Rufeger, U., Scharmann, W.: Comparative studies on isolated rat hepatocytes and AS-30 D hepatoma cells with leucocidin from Pseudomonas aeruginosa. Naunyn-Schmiedeberg's Arch. Pharmacol 295, 63–69 (1976)
Frimmer, M., Petzinger, E.: Mechanism of phalloidin intoxication. I: Cell membrane alterations. In: Membrane alterations as basis of liver injury (H. Popper, L. Bianchi, W. Reutter, eds.), pp. 293–299. Lancaster: MTP Press Ltd. 1977
Frimmer, M., Petzinger, E., Rufeger, U., Veil, L. B.: Trypsin protection of hepatocytes against phalloidin. Naunyn-Schmiedeberg's Arch. Pharmacol. 300, 163–171 (1977a)
Frimmer, M., Petzinger, E., Rufeger, U., Veil, L. B.: The role of bile acids in phalloidin poisoning. Naunyn-Schmiedeberg's Arch. Pharmacol. 301, 145–147 (1977b)
Govindan, V. M., Faulstich, H., Wieland, Th., Agostini, B., Hasselbach, W.: In vitro effect of phalloidin on a plasma membrane preparation of rat liver. Natuwissenschaften 59, 521–522 (1972)
Harris, H. E., Tso, M. Y. W., Epstein, H. F.: Actin and myosinlinked calcium regulation in the nematode Caenorhabditis elegans. Biochemical and structural properties of native filaments and purified proteins. Biochemistry 16, 859–865 (1977)
Kroker, R., Anwer, M. S., Hegner, D.: The lack of active bile acid transport in AS-30D ascites hepatoma cells. Naunyn-Schmiedeberg's Arch. Pharmacol. 303, 299–301 (1978)
Lengsfeld, A. M., Löw, J., Wieland, Th., Dancker, P., Hasselbach, W.: Interaction of phalloidin with actin. Proc. Natl. Acad. Sci. (U.S.A.) 71, 2803–2807 (1974)
Löw, J., Wieland, Th.: The interaction of phalloidin, some of its derivatives, and of other cyclic peptides with muscle actin as studied by viscosimetry. FEBS Lett. 44, 340–343 (1974)
Löw, J., Dancker, P., Wicland, Th.: Stabilization of actin induced by phalloidin at low pH. FEBS Lett. 65, 358–360 (1976)
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurements with the folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)
Michell, R. H., Hawthorne, J. N.: The site of diphosphoinositide synthesis in rat liver. Biochem. Biophys. Res. Commun. 21, 33 (1965)
Nemeth, E. F., Douglas, W. W.: Effects of microfilament-active drugs, phalloidin and the cytochalasins A and B, on exocytosis in mast cells evoked by 48/70 or A 23187. Naunyn-Schmiedeberg's Arch. Pharmacol. 302, 153–163 (1978)
Petzinger, E., Frimmer, M.: Studies on the phalloidin recognizing structure of the liver cell surface. Naunyn-Schmiedeberg's Arch. Pharmacol. 302, R18 (1978)
Petzinger, E., Seeger, R.: Scanning electron microscopic studies on the cytolytic effect of phallolysin on isolated rat hepatocytes and AS-30D hepatoma cells. Naunyn-Schmiedeberg's Arch. Pharmacol. 295, 211–213 (1976)
Petzinger, E., Homann, J., Frimmer, M.: Phalloidin-Antagonisten. 2. Mitteilung: Protektive Wirkung von Disilybin bei der Vergiftung isolierter Hepatozyten mit Phalloidin. Arzneim.-Forsch. (Drug Res.) 25, 571–576 (1975)
Petzinger, E., Grundmann, E., Veil, L. B., Frimmer, M., Fasold, H.: Inhibitory effect of 4,4′-diisothiocyano stilbene-2,2′-disulfonic acid (DIDS) in phalloidin response of isolated hepatocytes. Naunyn-Schmiedeberg's Arch. Pharmacol. 304, 303–307 (1978)
Reynolds, E. S.: The use of lead citrate at high pH as an electronopaque stain in electron microscopy. J. Cell Biol. 17, 208–211 (1963)
Rufeger, U., Grundmann, E.: Comparative studies on sensitivity to phalloidin of normal hepatocytes and AS-30D ascites hepatoma cells. Naunyn-Schmiedeberg's Arch. Pharmacol. 297, R17 (1977)
Schwarz, L. R., Burr, R., Schwenk, M., Pfaff, E., Greim, H.: Uptake of taurocholic acid into isolated rate-liver cells. Eur. J. Biochem. 55, 617–623 (1975)
Simionescu, N., Simionescu, M.: Galloylglucose of low molecular weight as mordant in electron microscopy. J. Cell Biol. 70, 608–621 (1976)
Wehland, J., Osborn, M., Weber, K.: Phalloidin-induced actin polymerization in the cytoplasm of cultured cells interferes with cell locomotion and growth. Proc. Natl. Acad. Sci. (U.S.A.) 74 5613–5617 (1977)
Wolff, J., Jones, A. B.: The purification of bovine thyroid plasma membranes and the properties of membrane-bound adenyl cyclase. J. Biol. Chem. 246, 3939–3947 (1971)
Wright, B. M., Edwards, A. J., Jones, V. E.: Use of a cell rupturing pump for the preparation of thymocyte subcellular fractions. J. Immunol. Methods 4, 281–286 (1974)
Author information
Authors and Affiliations
Additional information
This work was supported by the Deutsche Forschungsgemeinschaft
Rights and permissions
About this article
Cite this article
Grundmann, E., Petzinger, E., Frimmer, M. et al. A possible reason for the phalloidin tolerance of hepatoma cells. Naunyn-Schmiedeberg's Arch. Pharmacol. 305, 253–259 (1978). https://doi.org/10.1007/BF00498819
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00498819