Einleitung Für fortgeschrittene Tumore des Verdauungstraktes bestehen nur eingeschränkte Behandlungsoptionen. „Next generation sequencing“-Verfahren wie die gezielte Genpanelsequenzierung ermöglichen molekular getriebene Therapieansätze. Der Nutzen der Panelsequenzierung für die Therapie fortgeschrittener gastrointestinaler und hepatobiliärer Tumore ist unklar.
Lewis antigens and the Thomsen-Friedenreich (TF) antigen are complex glycan structures that modulate processes such as cell adhesion and proliferation and tumor metastasis. The aim of our study was to analyze the expression of sialyl Lewis A (sLeA), sialyl Lewis X (sLeX), Lewis Y (LeY), TF, galectin-1 (Gal-1) and galectin-3 (Gal-3) in human osteoblasts in vitro.The expression of the tumor markers sLeA, sLeX, LeY, TF, Gal-1 and Gal-3 was studied by means of immunohistochemistry on cells grown on chamber slides (2D) and on paraffin sections three-dimensional scaffold-free cultures (3D). The results of the stainings were evaluated semiquantitatively with the immunoreactive scoring system (IRS).Analysis of sLeA expression in both types of culture, 2D and 3D showed no detectable staining. After 5 days, in the 2D culture, expression of sLeX was weak, but the 3D culture (after 56 weeks) displayed a strong expression. LeY was expressed very slightly in the 2D culture, however LeY was not detectable in the 3D culture. The TF epitope was identified in the 2D cell culture model. In the 3D model, however, TF was completely lacking. Gal-1 was expressed very strongly in 2D culture, but in the 3D culture was not detectable. In contrast, Gal-3 was expressed in 3D culture but not in 2D.Within this study, we present a systematic analysis of the expression of sLeA, sLeX, LeY, TF, Gal-1 and Gal-3 in human osteoblasts grown in 2D and in 3D scaffold-free cultures. Summarizing the results of our study, we suggest that Lewis antigens and Gal-1 and -3 might play an important role in cell-cell and cell-matrix interactions of osteoblastic cells.
Trophoblast cells synthesize a variety of hormones, in which hCG plays a major role. On the strength of special enzymes they are capable of catalyzing the reaction cortisol <--> cortisone. In vitro experiments showed the influence on ACTH- and cortisol secretion by CRH, ACTH and prednisolon. In this study we describe the influence of cortisol (prednisolon) on hCG production of trophoblast cells in vitro.Trophoblast cells were prepared from human term placentae by standard trypsin-DNAse dispersion of villous tissue followed by a percoll gradient centrifugation step. After adjusting the cell suspension to a defined cell concentration of 1 x 10 (6) cells/ml cells were cultivated. The addition of prednisolon followed every eight hours. The samples were collected after 24 hours for a total of 96 hours also from unstimulated cultures. Culture supernatants were assayed for hCG by enzyme-immunometric methods.The addition of prednisolon (50 microg/ml) stimulates the concentration of hCG in a time-depending manner.The trophoblast cell shows an increase in the concentration of hCG after stimulation with cortisol. For the first time an influence of cortisol (prednisolon) on hCG production could be demonstrated in cultured trophoblast cells.
We review the current knowledge on alterations of the major basement membrane (BM) components and their cellular integrin receptors in benign and malignant tumors of epithelial and mesenchymal origin. While benign tumors usually exhibit a continous BM, recent analyses provide evidence that invasive growth of carcinomas coincides with (a) a loss in a proper BM, (b) changes in the type of integrin receptor expression and (c) the retained ability of certain tumor cells to synthesize matrix components. This latter aspect has been regarded as a potentially beneficial 'host' mechanism against invasive growth. This assumption is strongly supported by the finding of a positive correlation between the extent of BM loss and both a lesser degree of tumor differentiation and a worse prognosis of tumor growth. The resulting concept indicates that in carcinomas an imbalance in the cell-matrix interaction is the leading element in invasive growth. In mesenchymal tumors a somewhat different role of the BM can be observed. Thus, the qualitative and quantitative expression of major BM components in benign mesenchymal tumors closely relates to the BM pattern of normal tissues providing a histogenetically oriented classification of benign mesenchymal tumors. Most well-differentiated sarcomas retain a BM pattern close to that of the histogenetically related tissue, although in poorly differentiated sarcomas no such attribution to a histogenetic orientation of the tumor cells can be found.
Fragestellung: Therapeutische Antikörper gewinnen bei der Behandlung verschiedener Krebsarten mehr und mehr an Bedeutung. Ein Zielmolekül für eine antikörperbasierte Therapie ist Mucin 1 (MUC1). Epitheliale Tumore exprimieren Mucin 1 im Übermaß. Darüber hinaus fungiert Mucin 1 als Trägerprotein für onkofetale Kohlehydrat-Antigene wie Sialyl Lewis a und x, sowie Lewis y und dem Thomsen-Friedenreich Antigen. Der Antikörper Panko-Mab bindet spezifisch aktives Mucin 1, das noch membrangebunden und nicht abgekoppelt im Serum vorliegt. Methodik: Die Mammakarzinom-Zelllinien MCF7, T47D, CAMA, MDA-MB-435, ZR75–1 und MDA-MB-231 wurden in 96well-Platten für 48 Stunden mit steigenden Konzentrationen des anti-Muc1 Panko Mab inkubiert. Die Bestimmung der Zellproliferation erfolgte mittels Elisa über eine 5-Brom-2-desoxyuridin (BrdU)-Inkorporation, welches bei DNA-Neusynthese während der Mitose anstelle von Thymidin eingebaut wird.
