Zusammenfassung
Die Rolle der Tumorzell Interaktion mit der extrazellulären Matrix als auch die Tumorzell- Stromazell Interaktion sind für die Invasivität und somit für die Metastasierung von Tumorzellen von entscheidender Bedeutung. Experimentell kann die Tumorzell-Stromazell Interaktion zuverlässig mit Hilfe eines dreidimensionalen Konfrontationsassays, bei dem Tumorzellsphäroide mit embryonalen Hühnerherzsphäroiden konfrontiert werden, durchgeführt werden. Nach bereits 24h kann eine deutliche Infiltration von Tumorzellen in das embryonale Hühnerherzsphäroid mittels bildanalytischem Auswerteverfahren quantifiziert werden. Unter Einfluß von all-trans-Retinsäure (RA) findet eine Abnahme der Invasivität von Tumorzellen statt.
Sphäroide wurden auch verwendet, um die Tumorzell-Matrix Interaktion bezüglich gerichteter Zellmotilität unter Einfluß von RA zu untersuchen. Dabei wurden reaggregierte Tumor- bzw. Hühnerherzzellsphäroide auf verschiedenen Komponenten der extrazellulären Matrix und Komponenten der Basalmembran ausgebracht. Durch das Vermessen des sich nach 24h, 48h und 72h nach Ausbringung ergebenden Kreisdurchmessers, hervorgerufen durch auswandernde Zellen, kann eine Quantifizierung der gerichteten Motilität durchgeführt werden. Mit dieser in vitro-Methode zur Messung der gerichteten Motilität wurden Melanomzellen mit unterschiedlichem metastatischem Potential vom Menschen und von der Maus als auch embryonalen Hühnerherzzellen untersucht.
Es konnte durch diese Experimente gezeigt werden, daß die Geschwindigkeit mit der Zellen aus dem Sphäroid auswandern auf verschiedenen extrazellulären Matrixkomponenten unterschiedlich groß ist und daß die Invasivität nicht immer mit der gerichteten Motilität korreliert.
Unter Einfluß von lμM RA konnte auf Collagen I beschichtetem Glas bei den Mausmelanomzellen eine Zunahme des Kreisdurchmessers gegenüber den Kontrollwerten gemessen werden, wohingegen die embryonalen Hühnerherzzellen einen geringeren Kreisdurchmesser gegenüber den Kontrollwerten aufwiesen.
Summary
Tumour-host interaction and tumour interaction with the extracellular matrix plays a crucial role in invasiveness and determines cancer metastasis. To get insight in tumour-host interactions a reliable 3D confrontation assay could be performed where tumour spheroids will be confronted with chick embryonic heart spheroids. After 24h the infiltration of tumour cells into the chick embryonic heart spheroid can be evaluated using an image analysis Computer program. A reduction of tumour cell invasion can be seen under the influence of all-trans retinoic acid (RA).
Under the influence of RA spheroids were used to investigate the interaction of tumor cells with extracellular matrix concerning directional migration. Reaggregated spheroids consisting of tumour cells and chick embryonic heart cells were explanted on collagen or lamming coated surfaces. In evaluating the diameter change emerging from cells leaving the multicell spheroid after 24, 48 and 72 hours a quantification of directed motility can be made. We investigated cells with metastatic potential from human and murine origin and chick embryonic heart cells with this in vitro method of directional motility.
It could be shown that the directional migration depends on the coverslip coating and invasiveness is not strictly correlated with directional migration of the tumour cells. 1 μM all-trans Retinoic acid could increase the diameters in the case of murine melanoma cells whereas embryonic chick heart cells had decreased diameters on collagen Type I coated surfaces.
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Literatur
Albini A., Aukerman S.L., Ogle R . C., Noonan D.M. et al, The in vitro invasiveness and interactions with laminin of K-1735 melanoma cells. Evidence for different laminin-binding affmities in high and low metastatic variants, Clin Exp Metast, 7, 437 1989
Bertram J.S.,Role of gap junctional cell/cell communication in the control of proliferation and neoplastic transformation, Radiation Research, 123, 252–6, 1990
Boxberger H.-J. and Paweletz N.,Influences of various substrata on morphology, motility and invasiveness of rat tumour cells. II. Studies on the highly metastatic variant BSp 73 ASML, Anticancer Research, 10, 1265–74, 1990
Bracke M.E., Van Larebeke N.A., Vyncke B.M., Mareel M.M.,Retinoic acid modulates both invasion and plasma membrane ruffling of MCF-7 human mammary-carcinoma cells- in vitro, British Journal of Cancer, 63 (6), 867–72, 1991
Bräuner T.,Interzelluläre Kommunikation und invasives Wachstum maligner Zellen, in: Stuttgart: Verlag Stöffler & Schütz, 1987
Chelberg M.K., Tsilibary E.C., Hauser A . R., Mccarthy J.B.,Type IV Collagen-mediated melanoma cell adhesion and migration: involvement of multiple, distinct domains of the collagen molecule, Cancer Research, 49, 4796–802, 1989
Coopman P., Verhasselt B ., Bracke M., De-Bryne G. et al, Arrest of MCF-7 cell migration by laminin in vitro: possible mechanisms, Clinical Experimental Metastasis, 9, 460–84, 1991
Couchman J.R. and Rees D.A.,The behaviour of fibroblasts migrating from chick heart explants: changes in adhesion, locomotion and growth, and in the distribution of actomyosin and fibronectin, Journal of Cell Science, 39, 149–65, 1979
Engebraaten O., Schwachenwald R., Valen H., Bjerkvig R. et al, Effects o f high and l ow single dose irradiation on glioma spheroid invasion into normal rat brain tissue in vitro, Anticancer Research, 12, 1501–6, 1992
Fauennec L. and Cals M.J.,The biological effects of retinoids on cell differentiation and proliferation, Journal of Clinical Chemistry and Clinical Biochemistry, 26, 479–89, 1988
Fligiel S.E.G., Inman D.R., Talwar H.S., Fisher G.J. et al, Modulation of growth in normal and malignant melanocytic cells by all-trans retinoic acid, Journal of Cutaneous Pathology, 19, 27–33, 1992
Gehlsen K.R., Davis G.E., Sriramarao P., Integrin expression in human melanoma cells with deffering invasive and metastatic properties, Clinical Experimental Metastasis, 10, 111–20, 1992
Helige C., Smolle J., Zellnig G., Hartmann E. et al, Inhibition of K1735-M2 melanoma cell invasion in vitro by retinoic acid, Clin Exp Metast, 11, 409–18, 1993
Hendrix M.J.C., Wood W.R., Seftor E.A., Lotan D. et al., Retinoic acid inhibition of human melanoma cell invasion through a reconstituted basement membrane and its relation to decreases in the expression of proteolytic enzymes and motility factor receptor, Cancer Research, 50, 4121–30, 1990
Hofmann-Wellenhof R., Fink-Puches R., Smolle J., Helige C. et al., Correlation o f melanoma cell motility and invasion in vitro, Melanoma Research, 5, 311–9, 1995
Hossain M.Z. and Bertram J.S.,Retinoids suppress proliferation, induce cell spreading, and up-regulate connexin43 expression only in postconfluent 10T1/2 cells: implications for the role of gap junctional communication, Cell Growth and Differentiation, 5, 1253–61, 1994
Hossain M.Z., Wilkens L.R., Mehta P.P., Loewenstein W.R. et al., Enhancement o f gap junctional communication by retinoids correlates with their ability to inhibit neoplastic transformation, Carcinogenesis, 10, 1743–8,1989
Hossain M.Z., Zhang L.-X., Bertram J.,Progress in Cell Research 3: Gap junctions, in: Hall J., Zampighi G.A., Davis R.M. (eds.), London:Elsevier Science Publishers, 301–9, 1993
Kozlowski J.M., Hart I.R., Fidler I.J., Hanna N.,A human melanoma line heterogeneous with respect to metastatic capacity in athymic nude mice, Journal of the National Cancer Institute, 72, 913– 5, 1984
Lotan R.,Retinoids: New Trends in Research and Therapy, in: Saurat, Basel: Karger Verlag, 97–105, 1985
Lotan R., Amos B., Watanabe H., Raz A.,Suppression of Melanoma cell motility factor receptor expression by retinoic acid, Cancer Research, 52, 4878–84, 1992
Lotan R., Neumann G., Lotan D.,Modulation of cellular interactions by vitamin A and derivaties, in: De Luca L.M. and Shapiro S.S. (eds.), New York: New York Acad. Sei., 150–70, 1981
Mehta P.P., Bertram J.S., Loewenstein W.R.,The actions of retinoids on cellular growth correlate with their actions on gap junctional communication, Journal of Cell Biology, 108, 1053–65, 1989
Nakajima M., Lotan D., Baig M.M., Carralero R.M. et al., Inhibition by retinoic acid of type IV collagenolysis and invasion through reconstituted basement membrane by metastatic rat mammary adenocarcinoma cells, Cancer Research, 49, 1698–706, 1989
Pelzmann B.,Isolation und Kultivierung embryonaler Hühnerherzzellen, Diplomarbeit, 1990
Ranson M., Posen S., Mason R.S.,Extracellular matrix modulates the funetion of human melanocytes but not melanoma cells, Journal of Cellular Physiology, 136, 281–8, 1988
Silletti D. and Raz A.,Regulation of autoerine motility factor receptor expression in tumor cell locomotion and metastasis, Current Topical in Microbiology and Immunology, 213, 137–69, 1996
Storme G., Mareel M., De Bruyne G.,Influence of cell number on directional migration of M04 cells in vitro, Archiv für Geschwulstforschung, 51, 45–50, 1981
Storme G. and Mareel M.M.,Effects of anticancer agents on directional migration of malignant C3Hmouse fibroplastic cells in vitro, Cancer Research, 40, 943–8, 1980
Talmadge J.E. and Fidler I.J.,Enhanced metastatic potential of tumor cells harvested from spontaneous metastases of heterogeneous murine tumors, Journal of the National Cancer Institute, 69, 975–80, 1982
Tautt J., Mazur N., Koziorowska J.,Effects of retinoic acid on Plasminogen activator activity and cellular proliferation, Archivum Immunologiae et Therapiae Experimentalis, 38, 223–7,1990
Werling H.-O., Spiess E., Paweletz N., In vitro motility of BSp 73 rat tumor cells with different metastatic ability, Invasion and Metastasis, 6, 257–69, 1986
Wood W.R., Seftor E.A., Lotan D., Nakajima M. et al., Retinoic acid inhibits human melanoma tumor cell invasion, Anticancer Research, 10,423–32,1990
Woods A., Couchman J.R., Johansson S., Hoök M.,Adhesion and cytoskeletal Organisation of fibroblasts in response to fibronectin fragments, EMBO, 5,665–70, 1986
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Hartbauer, M., DeVaney, T., Ahammer, H., Tritthart, H. (2000). Die Motilität von menschlichen und Mausmelanomzellinien und embryonalen Hühnerherzzellen unter Einfluß von 1µM alltrans- Retinsäure. In: Schöffl, H., et al. Forschung ohne Tierversuche 2000. Ersatz- und Ergänzungsmethoden zu Tierversuchen. Springer, Vienna. https://doi.org/10.1007/978-3-7091-6760-1_23
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DOI: https://doi.org/10.1007/978-3-7091-6760-1_23
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