Zusammenfassung
Hintergrund: Tumorzell- und -gewebekulturen (2-dimensionale Monolayer [ML], 3-dimensionale multizelluläre
Tumorsphäroide [MTS] und Substrat-Kulturen [SK] auf Gelita) sollten biologische Eigenschaften
ihrer Ausgangstumoren repräsentieren. Kulturbedingungen u.a. können aber die Expression
zellulärer Antigene ändern. Methode: Ein Plattenepithelkarzinom des Hypopharynx wurde immunhistochemisch hinsichtlich
der Ausprägung der Zytokeratine (CK) 1, 7, 10, 14, 18 und 19, Vimentin, Neurofilamente
(NF) kD200 und kD68, Gangliosid CD2, Onkogen-Produkten (P53 mutiert und wild) und
membranassoziierten Antigenen (HLA-ABC und -DR, ECFR) an Gefrierschnitten (CS) und
an In-vitro-Kulturen (ML, MTS, SK) untersucht. Ergebnisse: Semiquantitativ waren deutliche Unterschiede in der Expression von CK1,14 und 19,
CD2 und P53 mutiert nachweisbar. MTS exprimierten die Marker mindestens genauso stark
wie die CS und stärker als die ML. Schlußfolgerungen: Die Antigen-Expression der MTS kam derjenigen in der Tumorbiopsie am nächsten. Die
schwache Expression in den SK kann durch lockere Zell-Zellkontakte bedingt sein.
Summary
Background: The need to improve therapy regims, determine prognosis, and study biological properties
of tumors extracorporally led to development of different experimental systems. In
order to approach the in vivo situation, specific properties of the tumors of origin
should be retained by the cells in culture over relatively long periods. However,
culture conditions may change expression of cellular antigens. Methods: Cryosections of a hypopharyngeal carcinoma were compared in this respect with different
cultivation systems (2-dimensional monolayers [ML], 3-dimensional multicellular tumor
spheroids [MTS] and substrate cultures on Gelita®) in regard to expression of cytokeratins
(CK) 1, 7, 10, 14, 18 and 19, vimentin, neurofilament (NF) kD200 and 68, ganglioside
GD2, oncogene products (P53 mutant and wild), and membrane-associated antigens (HLA-ABC
and -DR, epidermal growth factor receptor EGFR). Results: Semiquantitative immunohistochemical methods revealed differences in expression of
CK1,14 and 19, GD2, and P53 mutant between these systems. Conclusion: Pronounced expression of markers in MTS compared to original biopsy and monolayer
emphasizes the importance of cell-cell contact and 3-dimensionality or metabolic stress.
However, weak marker expression within substrate cultures may reflect loose cell-cell
contact observed. In these experiments, the antigenic configuration of MTS resembled the one of the
original tumor more than the other culture systems: monolayer and growth on substrate. As vimentin and NF are not expressed by healthy
epithelial cells of adults, occurrence of intratumoral vimentin and NF could point
to derepression of early differentiation antigens.
Schlüsselwörter
Plattenepithelkarzinome - In-vitro-Kulturen - Monolayer-Kulturen - Multizelluläre
Tumorsphäroide - Organkulturen - Intermediärfilamente - p53-Produkte - Membranassoziierte
Antigene - Immunhistochemie
Key words
Squamous cell carcinomas - In vitro culture - Monolayer cultures - Multicellular tumor
spheroids - Organ cultures - Intermediate filaments - P53 products - Membrane associated
antigens - Immunohistochemistry
