K Enderle-Ammour1,2, U Wellner3, E Kocsmar4, A Kiss4, G Lotz4, A Csanadi1,2, M Bader1, O Schilling5, M Werner1,6,2, P Bronsert7,8,9. 1. Institut für Klinische Pathologie, Universitätsklinikum Freiburg, Breisacher Straße 115A, 79106, Freiburg, Deutschland. 2. Medizinische Fakultät, Albert-Ludwigs-Universität Freiburg, Freiburg, Deutschland. 3. Klinik für Chirurgie, Universitätsklinikum Schleswig-Holstein, Campus Lübeck, Lübeck, Deutschland. 4. II. Institut für Pathologie, Semmelweis Universität, Budapest, Ungarn. 5. Institut für Molekulare Medizin und Zellforschung, Universitätsklinikum Freiburg, Freiburg, Deutschland. 6. Biobank Tumorzentrum Freiburg, Universitätsklinikum Freiburg, Freiburg, Deutschland. 7. Institut für Klinische Pathologie, Universitätsklinikum Freiburg, Breisacher Straße 115A, 79106, Freiburg, Deutschland. peter.bronsert@uniklinik-freiburg.de. 8. Biobank Tumorzentrum Freiburg, Universitätsklinikum Freiburg, Freiburg, Deutschland. peter.bronsert@uniklinik-freiburg.de. 9. Medizinische Fakultät, Albert-Ludwigs-Universität Freiburg, Freiburg, Deutschland. peter.bronsert@uniklinik-freiburg.de.
Abstract
BACKGROUND: In histopathological routine diagnostics, three-dimensional tissue samples are analyzed histologically and/or immunohistochemically in two-dimensional sectional planes due to the high expenditure of time and the lack of digitization possibilities. AIM: Here, we demonstrate the application of three-dimensional reconstruction to solid tumors and analyze inter-/intratumoral heterogeneity with respect to epithelial-mesenchymal transition (EMT). METHODS: Tissue samples from pancreatic, lung, colorectal, and breast cancers as well as colorectal liver metastases were serially processed in 4μm sections. For individual analyses, alternating stains (cytokeratin AE1/3, zinc finger E‑box-binding homeobox 1 (ZEB1), eCadherin) were performed. Subsequently, the tumor cells were analyzed for their morphology (epitheloid amoeboid, mesenchymal) and the expression of ZEB1 and eCadherin. For statistical analysis, all tumor cell aggregates were hierarchically annotated and analyzed. RESULTS: Tumor buds are predominantly associated with the main tumor mass. Furthermore, a shutteling of eCadherin could be observed within tumor cell aggregates smaller than nine cells. ZEB1 is only increasingly expressed in tumor cell groups smaller than five cells. CONCLUSIONS: The initial tumor budding and the subsequent decoupling of the tumor bud from the main tumor mass is most likely a two-part process. However, the EMT is not statistically significantly increased within the tumor bud detached from the main tumor mass. It could be shown that the currently valid and known definition of a tumor bud as a cell cluster of less than or equal to five cells cannot be completely classified in the concept of EMT represented by eCadherin and ZEB1.
BACKGROUND: In histopathological routine diagnostics, three-dimensional tissue samples are analyzed histologically and/or immunohistochemically in two-dimensional sectional planes due to the high expenditure of time and the lack of digitization possibilities. AIM: Here, we demonstrate the application of three-dimensional reconstruction to solid tumors and analyze inter-/intratumoral heterogeneity with respect to epithelial-mesenchymal transition (EMT). METHODS: Tissue samples from pancreatic, lung, colorectal, and breast cancers as well as colorectal liver metastases were serially processed in 4μm sections. For individual analyses, alternating stains (cytokeratin AE1/3, zinc finger E‑box-binding homeobox 1 (ZEB1), eCadherin) were performed. Subsequently, the tumor cells were analyzed for their morphology (epitheloid amoeboid, mesenchymal) and the expression of ZEB1 and eCadherin. For statistical analysis, all tumor cell aggregates were hierarchically annotated and analyzed. RESULTS:Tumor buds are predominantly associated with the main tumor mass. Furthermore, a shutteling of eCadherin could be observed within tumor cell aggregates smaller than nine cells. ZEB1 is only increasingly expressed in tumor cell groups smaller than five cells. CONCLUSIONS: The initial tumor budding and the subsequent decoupling of the tumor bud from the main tumor mass is most likely a two-part process. However, the EMT is not statistically significantly increased within the tumor bud detached from the main tumor mass. It could be shown that the currently valid and known definition of a tumor bud as a cell cluster of less than or equal to five cells cannot be completely classified in the concept of EMT represented by eCadherin and ZEB1.
Authors: E Sánchez-Tilló; A Lázaro; R Torrent; M Cuatrecasas; E C Vaquero; A Castells; P Engel; A Postigo Journal: Oncogene Date: 2010-04-26 Impact factor: 9.867
Authors: Ulrich Wellner; Jörg Schubert; Ulrike C Burk; Otto Schmalhofer; Feng Zhu; Annika Sonntag; Bettina Waldvogel; Corinne Vannier; Douglas Darling; Axel zur Hausen; Valerie G Brunton; Jennifer Morton; Owen Sansom; Julia Schüler; Marc P Stemmler; Christoph Herzberger; Ulrich Hopt; Tobias Keck; Simone Brabletz; Thomas Brabletz Journal: Nat Cell Biol Date: 2009-11-22 Impact factor: 28.824
Authors: Kirsten Aigner; Luise Descovich; Mario Mikula; Aneesa Sultan; Brigitta Dampier; Stefan Bonné; Frans van Roy; Wolfgang Mikulits; Martin Schreiber; Thomas Brabletz; Wolfgang Sommergruber; Norbert Schweifer; Andreas Wernitznig; Hartmut Beug; Roland Foisner; Andreas Eger Journal: FEBS Lett Date: 2007-03-21 Impact factor: 4.124