The effect of methotrexate on actin and invasiveness of hepatoma Morris 5123 cells in culture.

• Authors: Magdalena Otrocka , Hendrik Verschueren , Maria Malicka-Błaszkiewicz
• Languages of publication: EN

Abstracts

EN

Monomeric (G), total (T) and filamentous (F) actin and the state of actin polymerisation (F:G) were determined and actin filaments were visualized in hepatoma Morris 5123 cells cultured in the presence of methotrexate (MTX) at various concentration. The exposure of the cells to this drug resulted in a decrease of total and polymerised actin in cytoplasm and in some changes in actin filament organization. This coincided with a decrease of the cells' ability to migrate through Matrigel coated filters and with inhibition of tumour formation after reimplantation of the methotrexate treated cells to experimental rats.

Keywords

EN

hepatoma Morris 5123; invasiveness; methotrexate; actin

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2001
• Volume: 48
• Issue: 4
• Pages: 1051-1060

Dates

published

2001

received

2001-11-6

accepted

2001-12-4

Contributors

author

Magdalena Otrocka

• Department of Cell Pathology, Institute of Biochemistry and Molecular Biology, University of Wrocław, Wrocław, Poland

author

Hendrik Verschueren

• Pasteur Institut, Brussel, Belgium

author

Maria Malicka-Błaszkiewicz

• Department of Cell Pathology, Institute of Biochemistry and Molecular Biology, University of Wrocław, Wrocław, Poland

References

• 1. Janmey, P.A. & Chaponnier, C. (1995) Medical aspects of the actin cytoskeleton. Curr. Opin. Cell Biol. 7, 111-117.
• 2. Sheterline, P., Clayton, J. & Sparrow, J.C. (1998) Protein Profile. Actin, 4th edn. pp. 1-13, Oxford University Press, New York.
• 3. Ben-Ze’ev, A. (1985) The cytoskeleton in cancer cells. Biochim. Biophys. Acta 780, 197-212.
• 4. Hansell, E.J., Frisch, S.M., Tremble, P., Murnane, J.P. & Werb, Z. (1995) Simian virus 40 transformation alters the actin cytoskeleton, expression of matrix metalloproteinases and inhibitors of metalloproteinases, and invasive behavior of normal and ataxiatelagiectasia human skin fibroblasts. Biochem. Cell Biol. 73, 373-389.
• 5. Button, E., Shapland, C. & Lawson, D. (1995) Actin, its associated proteins and metastasis. Cell Motil. Cytoskeleton 30, 247-251.
• 6. Pokorna, E., Jordan, P.W., O’Neill, C.H., Zicha, D., Gilbert, C.S. & Vesely, P. (1994) Actin cytoskeleton and motility in rat sarcoma cell populations with different metastatic potential. Cell Motil. Cytoskeleton 28, 25-33.
• 7. Le, P.U., Nguyen, T.N., Drolet-Savoie, P., Leclerc, N. & Nabi, L.R. (1998) Increased β-actin expression in an invasive Moloney sarcoma virus-transformed MDCK cell variant concentrates to the tips of multiple pseudopodia. Cancer Res. 58, 1631-1635.
• 8. Suzuki, H., Nagata, H., Shimada, Y. & Konno, A. (1998) Decrease in γ-actin expression, disruption of actin filaments and alterations in cell adhesion in human salivary gland adenocarcinoma cell clones. Int. J. Oncol. 12, 1079-1084.
• 9. Helige, C., Zellnig, G., Hofmann-Wellenhof, R., Fink-Puches, R., Smolle, J. & Tritthart, H.A. (1997) Interrelation of motility, cytoskeletal organization and gap junctional communication with invasiveness of melanocytic cells in vitro. Invasion Metastasis 17, 26-41.
• 10. Verschueren, H., Van Der Taelen, I., Dewit, J., De Braekeleer, J. & De Beatselier, P. (1994) Metastatic competence of BW5147 T-lymphoma cell lines is correlated with in vitro invasiveness, motility and F-actin content. J. Leukoc. Biol. 55, 552-556.
• 11. Verschueren, H., Van Der Taelen, I., Dewit, J., De Braekeleer, J., De Beatselier, P., Aktories, K. & Just, I. (1995) Effects of Clostridium botulinum C2 toxin and cytochalasin D on in vitro invasiveness, motility and F-actin content of a murine T-lymphoma cell line. Eur. J. Cell Biol. 66, 335-341.
• 12. Iwazaki, R., Watanabe, S., Otaka, K., Ota, K., Ono, Y. & Sato, N. (1997) The role of the cytoskeleton in migration and proliferation of a cultured human gastric cancer cell line using a new metastasis model. Cancer Lett. 119, 191-196.
• 13. Stournaras, C., Stiakaki, E., Koukouritaki, S.B., Theodoropoulos, P.A., Kalmanti, M., Fostinis, Y. & Gravanis, A. (1996) Altered actin polymerisation dynamics in various malignant cell types: Evidence for differential sensitivity to cytochalasin B. Biochem. Pharmacol. 52, 1339-1346.
• 14. Malicka-Błaszkiewicz, M., Styczeń, I., Nowak, D., Hańczycowa, H., Ponikowski, P. & Sebzda, T. (1995) Actin content and polymerisation in tumour, liver and serum of the hepatoma Morris 5123 tumour bearing rats. Mater Med. Pol. 27, 115-118.
• 15. Nowak, D., Majcher, I., Kochman, A. & Malicka-Blaszkiewicz, M. (1995) The changes in actin content and polymerisation during hepatoma Morris 5123 tumour development. J. Exp. Clin. Cancer Res. 14, 37-40.
• 16. Jordan, M.A. & Wilson, L. (1998) Microtubules and actin filaments: Dynamic targets for cancer chemotherapy. Curr. Opin. Cell Biol. 10, 123-130.
• 17. Bubb, M.R., Spector, I., Bershadsky, A.D. & Korn, E.D. (1995) Swinholide A is a microfilament disrupting marine toxin that stabilizes actin dimers and severs actin filaments. J. Biol. Chem. 270, 3463-3466.
• 18. Suenaga, K., Kamei, N., Okugawa, Y., Takagi, M., Akao, A., Kigoshi, H. & Yamada, K. (1997) Cytotoxicity and actin depolymerizing activity of aplyronine A, a potent antitumour macrolide of marine origin, and the natural and artificial analogs. Bioorg. Med. Chem. Lett. 7, 269-274
• 19. Jolivet, J., Cowan, K.H., Curt, G.A., Clendeninn, N.J. & Chabner, B.A. (1983) The pharmacology and clinical use of methotrexate. N. Engl. J. Med. 309, 1094-1104.
• 20. Takemura, Y. & Jackman, A.L. (1997) Folate-based thymidylate synthase inhibitors in cancer chemotherapy. Anticancer Drugs 8, 3-16.
• 21. Laurance, J.E. & Sandborn, W.J. (1996) Methotrexate for inflammatory bowel disease: Pharmacology and preliminary results. Mayo Clin. Proc. 71, 69-80.
• 22. Otrocka, M., Nowak, D., Sobczak, I. & Malicka-Błaszkiewicz, M. (1999) The effect of methotrexate (MTX) on the changes in actin content and polymerisation during hepatoma Morris 5123 growth process. Diag. Lab. 35, 399-407.
• 23. Morris, H.P., Sidransky, H., Wagner, B.P. & Dyer, H.M. (1960) Some characteristic of transplantable rat hepatoma 5123 induced by ingestion of N-(2-fluorenyl)phthalamic acid. Cancer Res. 20, 1252-1254.
• 24. Doyle, A., Griffiths, J.B. & Newell, D.G. (1993) Basic techniques for primary cell cultures and establishment of continuous cultures; in: Cell and Tissue Culture: Laboratory Procedures, chap. 3E, pp. 2.1-2.8, John Wiley & Sons, Chichester.
• 25. Albini, A., Iwamoto, Y., Kleinman, H.K., Martin, G.R., Aaronson, S.A., Kozlowski, J.M. & Mcewan, R.N. (1987) A rapid in vitro assay for quantitating invasive potential of tumour cells. Cancer Res. 47, 3239-3245.
• 26. Malicka-Błaszkiewicz, M. & Roth, J.S. (1981) Some factors affecting the interaction between actin leukemia L1210 cells and DNase I. Biochem. Biophys. Res. Commun. 102, 594-601.
• 27. Malicka-Błaszkiewicz, M. (1986) DNase-like activity and actin content in the liver of some vertebrates. Comp. Biochem. Physiol. 84B, 207-209.
• 28. Lowry, O.H., Rosebrough, N.J., Farr, A.L & Randall, R.J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265-275.
• 29. Bond, M. & Somlyo, A.V. (1982) Dense bodies and actin polarity in vertebrate smooth muscle. J. Cell Biol. 95, 403-413.
• 30. Duncan, K., Duncan, M., Alley & M. Sausville, E. (1996) Cucurbitacin E-induced disruption of the actin and vimentin cytoskeleton in prostate carcinoma cells. Biochem. Pharmacol. 52, 1553-1560.
• 31. Zeng, H., Xu, Z. & Wang, K. (1997) FT-Raman studies on the transformation of G-actin to F-actin, the binding of cisplatin and transplantin to F-actin and the effect of conformation of F-actin. Int. J. Biol. Macromol. 20, 107-113.
• 32. Kruidering, M., van de Water, B., Zhan, Y., Baelde, J.J., de Heer, E., Mulder, G.J., Stevens, J.L. & Nagelkerke, J.F. (1998) Cisplatin effects on F-actin and matrix proteins precede renal tubular cell detachment and apoptosis in vitro. Cell Death Differ. 5, 601-614.
• 33. Rosenblum, M. & Shivers, R. (2000) ‘Rings’ of F-actin form around the nucleus in cultured human MCF7 adenocarcinoma cells upon exposure to both taxol and taxotere. Comp. Biochem. Physiol. 125, 121-131.
• 34. Tikoo, A., Shakri, R., Connolly, L., Hirokawa, Y., Shishido, T., Bowers, B., Ye, L., Kohama, K., Simpson, R.J. & Maruta, H. (2000) Treatment of ras-induced cancers by the F-actin-bundling drug MKT-077. Cancer J. Sci. Am. 6(3), 162-168.
• 35. Terzis, A.J.A., Fiskerstrand, T., Refsum, H., Ueland, P.M., Arnold, H. & Bjerkvig, R. (1993) Proliferation, migration and invasion of human glioma cells exposed to antifolate drugs. Int. J. Cancer 54, 112-118.