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

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Otrocka M., Verschueren H., Malicka-Błaszkiewicz M.
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2001
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vol. 48
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issue 4
1051-1060
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.
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Identification of actin from hepatoma Morris 5123 cells

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Nowak D., Kochman A., Malicka-Błaszkiewicz M.
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EN
The hepatoma Morris 5123 tumor growth is accompanied by changes in actin content and polymerization (Malicka-Błaszkiewicz et al. (1995) Mat. Med. Pol., 27, 115-118;  Nowak et al. (1995) J. Exp. Cancer Res. 14, 37-40). Presently actin isoforms from cytosol and  cytoskeleton fractions were separated by SDS/PAGE and identified with antibodies directed against different actin isoforms. Actin isolated from the cytosol by affinity chromatography on DNase I bound to agarose shows the presence of only one protein spot on 2D gel electrophoresis corresponding to the mobility of the rabbit α skeletal muscle actin (M^r 43000) and isoelectric point equal to 5.3. It interacts only with monoclonal anti β actin isoform antibodies, posing the question of differential affinity of actin isoforms to DNase I.
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