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Three-dimensional analysis of the substrate-dependent invasive behavior of a human lung tumor cell line with a confocal laser scanning microscope

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Abstract

Matrigel and collagen G gels were used as models for basement membrane and interstitial space-collagen, respectively, to study the invasive behavior of cells of the human lung tumor cell line EPLC 32M1, which was derived from a squamous cell carcinoma. For three dimensional analysis of the invasive process, cells were seeded onto the gels in a slide chamber and observed with a confocal laser scanning microscope. Optical sectioning in thexy andxz directions and image reconstruction with computer programs allowed us readily to obtain a three-dimensional overview of the invasive process in situ. Both types of gel showed a smooth surface. Matrigel had a granular structure whereas collagen G revealed a fiber-like morphology. The tumor cells showed a matrix-dependent behavior. On Matrigel, within 24 h of incubation, a network of cells appeared on the surface, which developed further within 72 h to interconnected multicellular cords also invading the gel. Tumor cells seeded on collagen G remained individual. They formed pseudopodia and achieved tight contact with the matrix, eventually also invading the gels in a time-dependent manner. Therefore, the composition of the substrate crucially influences the invasion path.

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References

  • Albini A, Iwamoto Y, Kleinman HK, Martin GR, Aaronson SA, Kozlowski JM, McEwan RN (1987) A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res 47:3239–3245

    PubMed  Google Scholar 

  • Azuma M, Tamatani T, Fukui K, Bando T, Sato M (1994) Enhanced proteolytic activity is responsible for the aberrant morphogenetic development of SV40-immortalized normal human salivary gland cells grown on basement membrane components. Lab Invest 70:217–227

    PubMed  Google Scholar 

  • Bepler G, Köhler A, Kiefer P, Havemann K, Beisenherz K, Jaques G, Gropp C, Haeder M (1988) Characterisation of the state of differentiation of six newly established human non-small-cell lung cancer cell lines. Differentiation 37:158–171

    PubMed  Google Scholar 

  • Boxberger H-J, Paweletz N, Spiess E, Kriehuber R (1989) An in vitro study of Bsp73 rat tumor cell invasion into endothelial monolayer. Anticancer Res 9:1777–1786

    PubMed  Google Scholar 

  • Heidtmann HH, Hofmann M, Jacob E, Erbil C, Havemann K, Schwartz-Albiez R (1989) Synthesis and secretion of plasminogen activators and plasminogen activator inhibitors in cell lines of different groups of human lung tumors. Cancer Res 49:6960–6965

    PubMed  Google Scholar 

  • Heidtmann HH, Elsässer HP, Salge U, Hejmanns J, Erbil C, Schwartz-Albiez R, Havemann K (1992) Plasminogen activator system and xenograft growth patterns in human non-small-cell lung cancer cell lines. Fibrinolysis 6 [Suppl 4]:77–82

    Google Scholar 

  • Imamura H, Takao S, Aikou T (1994) A modified invasion-3-(4,5-dimenthylthiazole-2-yl)-2,5-diphenyltetrazolium bromide assay for quantitating tumor cell invasion. Cancer Res 54:3620–3624

    PubMed  Google Scholar 

  • Inoué S, Leblond CP, Laurie GW (1983) Ultrastructure of Reichert's membrane, a multilayered basement membrane on the parietal wall of the rat yolk sac. J Cell Biol 97:1524–1537

    PubMed  Google Scholar 

  • Kleinman HK, McGarvey ML, Hassel JR, Star VL, Cannon FB, Laurie GW, Martin GR (1986) Basement membrane complexes with biological activity. Biochemistry 25:312–318

    PubMed  Google Scholar 

  • Kramer RH, Bensch KG, Wong J (1986) Invasion of reconstituted basement membrane matrix by metastatic human tumor cells. Cancer Res 46:1980–1989

    PubMed  Google Scholar 

  • Kubota Y, Kleinman HK, Martin GR, Lawley TJ (1988) Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures. J Cell Biol 107:1589–1598

    PubMed  Google Scholar 

  • Kuzuya M, Kinsella JL (1994) Induction of endothelial cell differentiation in vitro by fibroblast-derived soluble factors. Exp Cell Res 215:310–318

    PubMed  Google Scholar 

  • Lee J, Ishihara A, Jacobson K (1993) How do cells move along surfaces? Trends Cell Biol 3:366–370

    PubMed  Google Scholar 

  • Liotta LA (1986) Tumor invasion and metastases: role of extracellular matrix. Rhoads Memorial Award Lecture. Cancer Res 46:1–7

    PubMed  Google Scholar 

  • Liotta LA, Lee WC, Morakis DJ (1980) New method for preparing large surfaces of intact basement membrane for tumor invasion studies. Cancer Lett 11:141–152

    PubMed  Google Scholar 

  • Madri JA, Williams SK, Wyatt T, Mezzio C (1983) Capillary endothelial cell cultures: phenotypic modulation by matrix components. J Cell Biol 97:153–165

    PubMed  Google Scholar 

  • Martin GR, Timpl R (1987) Laminin and other basement membrane components. Annu Rev Cell Biol 3:57–85

    PubMed  Google Scholar 

  • Mignatti P, Rifkin DB (1993) Biology and biochemistry of proteinases in tumor invasion. Physiol Rev 73:161–195

    PubMed  Google Scholar 

  • Montcourrier P, Mangeat PH, Valembois C, Salazar G, Sahuquet A, Duperray C, Rochefort H (1994) Characterization of very acidic phagosomes in breast cancer cells and their association with invasion. J Cell Sci 107:2381–2391

    PubMed  Google Scholar 

  • Noël AC, Callé A, Emonard HP, Nusgens BV, Simar L, Foidart J, Lapiere CM, Foidart J-M (1991) Invasion of reconstituted basement membrane matrix is not correlated to the malignant metastatic cell phenotype. Cancer Res 51:405–414

    PubMed  Google Scholar 

  • Simon N, Noël A, Foidart J-M (1992) Evaluation of in vitro reconstituted basement membrane assay to assess the invasiveness of tumor cells. Invasion Metastasis 12:156–167

    PubMed  Google Scholar 

  • Sloane BF, Moin K, Krepela E, Rozhin J (1990) Cathepsin B and its endogenous inhibitors: the role in tumor malignancy. Cancer Metastasis Rev 9:333–352

    PubMed  Google Scholar 

  • Spanel-Borowski K, Ricken A, Patton WF (1994) Cytokeratin-positive and cytokeratin-negative cultured endothelial cells from bovine aorta and vena cava. Differentiation 57:225–234

    PubMed  Google Scholar 

  • Spring H, Franke WW (1981) Transcriptionally active chromatin in loops of lampbrush chromosomes at physiological salt concentrations as revealed by electron microscopy of sections. Eur J Cell Biol 24:298–308

    PubMed  Google Scholar 

  • Stetler-Stevenson WG, Liotta LA, Kleiner DE (1993) Extracellular matrix: role of matrix metalloproteinase in tumor invasion and metastasis. FASEB J 7:1434–1441

    PubMed  Google Scholar 

  • Ulbricht B, Spiess E, Schwartz-Albiez R, Ebert W (1995) Quantification of intracellular cathepsin activities in human lung tumor cell lines by flow cytometry. Biol Chem. Hoppe Seyler 376:407–414

    PubMed  Google Scholar 

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Strohmaier, AR., Spring, H. & Spiess, E. Three-dimensional analysis of the substrate-dependent invasive behavior of a human lung tumor cell line with a confocal laser scanning microscope. Histochem Cell Biol 105, 179–185 (1996). https://doi.org/10.1007/BF01462290

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