Beta-actin in human colon adenocarcinoma cell lines with different metastatic potential

• Autorzy: Nowak D. , Skwarek-Maruszewska A. , Zemanek-Zboch M. , Malicka-Blaszkiewicz M.
• Języki publikacji: EN

Abstrakty

EN

Human colon adenocarcinoma LS180 parental cell line and selected variants, characterized by different metastatic capacity were used to examine, whether a correlation exists between β-actin expression, its subcellular distribution and metastatic potential of these cells. Cytosolic fraction (supernatant 105 000 × g), isolated from the tumor cells was used as a source for actin quantification. The higher level of β-actin was observed in the cytosol of three selected sublines to compare with LS180 parental line. Statistically significant increase of β-actin level in highly motile EB3 cells variant should be underlined to compare with the other sublines. Distinct differences in the phenotype of adenocarcinoma cell variants were found, such as the changes in cells shape, cells spreading and ability to attach to the surface of culture dish. Actin cytoskeleton was visualized with fluorescence microscopy application and microfilaments rhodamine-conjugated phalloidin staining. β-actin subcellular localization was done by immunofluorescence staining with monoclonal anti-β actin antibodies. In the elongated cells (LS180, 3LNLN), this isoactin is dispersed in the whole cell body and concentrates in pseudopods and at the leading edges, when in the rounded variant (EB3) β-actin dominates mainly in cortical ring under cellular membrane and it is also seen in the subtle protrusions. Summary of our former (Nowak et al., 2002, Acta Biochim. Polon., 49: 823) and current data lead to the conclusion that there is a distinct correlation between metastatic capacity of examined human colon adenocarcinoma cells, the state of actin polymerization, actin cytoskeleton organization and β-actin expression.

Słowa kluczowe

EN

cell line; beta-actin; man; adenocarcinoma; invasiveness; actin cytoskeleton; colon adenocarcinoma

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2005
• Tom: 52
• Numer: 2
• Opis fizyczny: p.461-468,fig.,ref.

Twórcy

autor

Nowak D.

• University of Wroclaw, Wroclaw, Poland

autor

Skwarek-Maruszewska A.

autor

Zemanek-Zboch M.

autor

Malicka-Blaszkiewicz M.

Bibliografia

• Bond M, Somlyo AV (1982) Dense bodies and actin polarity in vertebrate smooth muscle. J Cell Biol 95: 403–413.
• Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing, the principle of protein-dye binding. Anal Biochem 272: 248–254.
• Buda A, Pignatelli M (2004) Cytoskeletal network in colon cancer: from genes to clinical application. Int J Biochem 36: 759–765.
• Button E, Shapland C, Lawson D (1995) Actin, its associated proteins and metastasis. Cell Motil Cytoskel 30: 247–251.
• Chaponnier C, Gabbiani J (2004) Pathological situations characterized by altered isoform expression. J Pathol 204: 386–395.
• Herman IM (1993) Actin isoforms. Curr Opin Cell Biol 5: 48–55.
• Honda K, Yamada T, Hayashida Y, Idogawa M, Sato S, Hasegawa F, Ino Y, Ono M, Hirohashi S (2005) Actinin- 4 increases cell motility and promotes lymph node metastasis of colorectal cancer. Gastroenterology 128: 51–62.
• Hood JD, Cheresh DA (2002) Role of integrins in cell invasion and migration. Nat Rev 2: 91–100.
• Janmey PA, Chaponnier C (1995) Medical aspects of the actin cytoskeleton. Curr Opin Cell Biol 7: 111–117.
• Kassis J, Lauffenburger DA, Turner T, Wells A (2001) Tumour invasion as dysregulated cell motility. Cancer Biol 11: 105–117.
• Keller HU, Bebie H (1996) Protrusive activity quantitatively determines the rate and direction of cell locomotion. Cell Motil Cytoskel 33: 241–251.
• Keller H, Eggli P (1998) Actin accumulation in pseudopods or in the tail of polarized walker carcinosarcoma cells quantitatively correlates with local folding of the cell surface membrane. Cell Motil Cytoskel 40: 342–353.
• Khatilina S (2001) Functional specificity of actin isoforms. Inter Rev Cytol 202: 35–98.
• Kieda C, Paprocka M, Krawczenko A, Załęcki P, Dupuis P, Monsigny M, Radzikowski C, Duś D (2002) New human microvascular endothelial cell lines with specific adhesion molecules phenotypes. Endothelium 9: 247–261.
• Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
• Lambrechts A, Van Troys M, Ampe C (2004) The actin cytoskeleton in normal and pathological motility. Int J Bio em Cell Biol 36: 1890–1909.
• Le PU, Nguyen TN, Drolet-Savoie P, Leclerc N, Nabi LR (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.
• Malicka-Błaszkiewicz M, Roth JS (1981) Some factors affecting the interaction between actin leukemia L1210 cells and DNase I. Biochem Biophys Res Commun 102: 594–601.
• Malicka-Błaszkiewicz M, Styczeń I, Nowak D, Hańczycowa H, Ponikowski P, Sebzda T (1995) Actin content and polymerization in tumor, liver and serum of the hepatoma Morris 5123 tumor bearing rats. Mat Med Pol 27: 115–118.
• Nagata K, Ichikawa Y (1984) Changes in actin during differentiation. Cell Muscle Motil 5: 171–193.
• Nguyen TN, Wang HJ, Zalzal S, Nanci A, Nabi IR (2000) Purfication and characterization of β actin-rich tumor cell pseudopodia: role of glycolysis. Exp Cell Res 258: 171–183.
• Nowak D, Majcher I, Kochman A, Malicka-Błaszkiewicz M (1995) The changes in actin content and polymerization during hepatoma Morris 5123 tumor development. J Exp Clin Cancer Res 14: 37–40.
• Nowak D, Kochman A, Malicka-Błaszkiewicz M (1999) Identification of actin from hepatoma Morris 5123 cells. Acta Biochim Polon 46: 949–959.
• Nowak D, Malicka-Błaszkiewicz M (1999) Izoformy aktyny– zrożnicowanie funkcji, zmiany w stanach patologicznych. Post Biochem 45: 261–269 (in Polish).
• Nowak D, Krawczenko A, Duś D, Malicka-Błaszkiewicz M (2002) Actin in human colon adenocarcinoma cells with different metastatic potential. Acta Biochim Polon 49: 823–828.
• Opolski A, Wietrzyk J, Duś D, Kieda C, Matejuk A, Makowska A, Wojdat E, Ugorski M, Laskowska A, Kłopocki A, Rygaard J, Radzikowski C (1998) Metastatic potential and saccharide antigens expression of human colon cancer cells xenotransplanted into athymic nude mice. Folia Microbiol 43: 507–510.
• Otrocka M, Verschueren H, Malicka-Błaszkiewicz M (2001) The effect of methotrexate on actin and invasiveness of hepatoma Morris 5123 cells in culture. Acta Biochim Polon 48: 1051–1060.
• Peckham M, Miller G, Wells C, Zicha D, Dunn GA (2001) Specific changes to the mechanism of cell locomotion induced by overexpression of β actin. J Cell Sci 114: 1367–1377.
• Pokorna E, Jordan PW, O’Neill CH, Zicha D, Gilbert CS, Vesely P (1994) Actin cytoskeleton and motility in rat sarcoma cell populations with different metastatic potential. Cell Motil Cytoskel 28: 25–33.
• Rędowicz MJ (2002) Myosins and pathology: genetics and biology. Acta Biochim Polon 49: 789–804.
• Sahai E, Marshall CJ (2003) Differing modes of tumour cell invasion have distinct requirements for Rho/ROCK signaling and extracellular proteolysis. Nat Cell Biol 5: 711–719.
• Shestakova EA, Wyckoff J, Jones J, Singer RH, Condeelis J (1999) Correlation of β-actin messenger RNA localization with metastatic potential in rat adenocarcinoma cell lines. Cancer Res 59: 1202–1205.
• Sheterline P, Clayton J, Sparrow JC (1998) Protein Profile. Actin, Oxford University Press, New York.
• Stournaras C, Stiakaki E, Koukouritaki SB, Theodoropoulos PA, Kalmanti M, Fostinis Y, Gravanis A (1996) Altered actin polymerization dynamics in various malignant cell types: evidence for differential sensitivity to cytochalasin B. Biochem Pharmacol 52: 1339–1346.
• 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.
• Towbin H, Stachelin T, Gordon T (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 466–472.
• Vandekerckhove J, Weber K (1978) At least six different actins are expressed in a higher mammal: An analysis based on the aminoacids sequence of the amino-terminal tryptic peptide. J Biol Chem 126: 783–802.
• 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.
• Welch AY, Herman IM (2002) Cloning and characterization of βCAP73, a novel regulator of β actin assembly. Int J Biochem Cell Biol 34: 864–881.
• Wong K, Rubenthiran U, Jothy S (2003) Motility of colon cancer cells: modulation by CD44 isoform expression. Exp Mol Pathol 75: 124–130.
• Wyckhoff JB, Jones JG, Condeelis JS, Segall JE (2000) A critical step in metastasis: in vitro analysis of intravasation at the primary tumor. Cancer Res 60: 2504–2511.