caa a22 4500 475773969 CHVBK 20180406123625.0 cr unu---uuuuu 170329e20000701xx s 000 0 eng 10.1007/s004180000159 doi (NATIONALLICENCE)springer-10.1007/s004180000159 Simultaneous enzyme overlay membrane (EOM)-based in situ zymography and immunofluorescence technique reveals cathepsinB-like activity in a subset of tumour vessels [Elektronische Daten] [Alexander Berndt, Jördis Johannesson, Michael Haas, Christoph Arkona, Detlef Katenkamp, Bernd Wiederanders, Peter Hyckel, Hartwig Kosmehl] Abstract.: The analysis of the invasion front of oral squamous cell carcinoma (OSCC) has revealed a fundamental invasion-associated remodelling of the extracellular matrix as the result of a complex regulated interplay of matrix synthesis and matrix degradation. Cysteine proteinases, in particular cathepsinB, are implicated in tumour invasion in vivo and in vitro and are thought to be important mediators of metastasis. An in situ zymographic assay based on enzyme overlay membranes (EOMs) was established to define the tissue localisation of cathepsinB activity in OSCC. Using confocal laser scanning microscopy we present a double-labelling method for the rapid and reproducible simultaneous detection of cathepsinB-like activity and cellular or extracellular antigens based on an EOM and immunofluorescence technique on frozen sections. Applying this method, cathepsinB-like activity was mainly found in vascular structures within the invasive front of OSCC. Therefore, the results suggest a particular pathogenic role of cathepsinB in tumour angioneogenesis. The method can simply be transferred to other enzymes and can be recommended for more extensive studies of proteolytic activity in human malignancies. Springer-Verlag, 2000 Angioneogenesis CathepsinB Enzyme overlay membrane In situ zymography Oral squamous cell carcinoma nationallicence Berndt Alexander Institute of Pathology, Friedrich Schiller University, 07740 Jena, Germany aut Johannesson Jördis Institute of Pathology, Friedrich Schiller University, 07740 Jena, Germany aut Haas Michael Institute of Pathology, Friedrich Schiller University, 07740 Jena, Germany aut Arkona Christoph Institute of Biochemistry, Friedrich Schiller University Jena, Germany aut Katenkamp Detlef Institute of Pathology, Friedrich Schiller University, 07740 Jena, Germany aut Wiederanders Bernd Institute of Biochemistry, Friedrich Schiller University Jena, Germany aut Hyckel Peter Clinic of Maxillofacial Surgery/Plastic Surgery, Friedrich Schiller University Jena, Germany aut Kosmehl Hartwig Institute of Pathology, Friedrich Schiller University, 07740 Jena, Germany aut https://doi.org/10.1007/s004180000159 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s004180000159 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Berndt Alexander Institute of Pathology, Friedrich Schiller University, 07740 Jena, Germany aut NATIONALLICENCE 700 1- Johannesson Jördis Institute of Pathology, Friedrich Schiller University, 07740 Jena, Germany aut NATIONALLICENCE 700 1- Haas Michael Institute of Pathology, Friedrich Schiller University, 07740 Jena, Germany aut NATIONALLICENCE 700 1- Arkona Christoph Institute of Biochemistry, Friedrich Schiller University Jena, Germany aut NATIONALLICENCE 700 1- Katenkamp Detlef Institute of Pathology, Friedrich Schiller University, 07740 Jena, Germany aut NATIONALLICENCE 700 1- Wiederanders Bernd Institute of Biochemistry, Friedrich Schiller University Jena, Germany aut NATIONALLICENCE 700 1- Hyckel Peter Clinic of Maxillofacial Surgery/Plastic Surgery, Friedrich Schiller University Jena, Germany aut NATIONALLICENCE 700 1- Kosmehl Hartwig Institute of Pathology, Friedrich Schiller University, 07740 Jena, Germany aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer