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Int. J. Mol. Sci. 2016, 17(10), 1655; doi:10.3390/ijms17101655

Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines

1
Small Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559 Hannover, Germany
2
Division of Medicine, Haematology, Oncology and Palliative Medicine, University of Rostock, Ernst-Heydemann-Str. 6, 18055 Rostock, Germany
3
Institute of Biophysics, Leibniz University Hannover, Herrenhäuser Straße 2, 30419 Hannover, Germany
4
Institute for Bioinformatics, University Medicine Greifswald, Walther-Rathenau-Str. 48, 17475 Greifswald, Germany
5
Institute of Veterinary Medicine, Georg-August-University Göttingen, Burckhardtweg 2, 37077 Göttingen, Germany
6
Department of Pathology, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany
7
Center for Systems Neuroscience (ZSN) Hannover, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany
*
Author to whom correspondence should be addressed.
Academic Editors: Ge Zhang, Aiping Lu and Hailong Zhu
Received: 11 August 2016 / Revised: 20 September 2016 / Accepted: 21 September 2016 / Published: 29 September 2016
(This article belongs to the Special Issue Translational Molecular Medicine & Molecular Drug Discovery)
View Full-Text   |   Download PDF [3019 KB, uploaded 29 September 2016]   |  

Abstract

Human and canine mammary tumours show partial claudin expression deregulations. Further, claudins have been used for directed therapeutic approaches. However, the development of claudin targeting approaches requires stable claudin expressing cell lines. This study reports the establishment and characterisation of canine mammary tissue derived cell lines, analysing longitudinally the claudin-1, -3, -4 and -7 expressions in original tissue samples, primary cultures and developed cell lines. Primary cultures were derived from 17 canine mammary tissues: healthy, lobular hyperplasia, simple adenoma, complex adenoma, simple tubular carcinoma, complex carcinoma, carcinoma arising in a benign mixed tumour and benign mixed tissue. Cultivation was performed, if possible, until passage 30. Claudin mRNA and protein expressions were analysed by PCR, QuantiGene Plex Assay, immunocytochemistry and immunofluorescence. Further, cytokeratin expression was analysed immunocytochemically. Cultivation resulted in 11 established cell lines, eight showing epithelial character. In five of the early passages the claudin expressions decreased compared to the original tissues. In general, claudin expressions were diminished during cultivation. Three cell lines kept longitudinally claudin, as well as epithelial marker expressions, representing valuable tools for the development of claudin targeted anti-tumour therapies. View Full-Text
Keywords: claudin; mammary neoplasias; canine; cell lines; cell culture; marker expression claudin; mammary neoplasias; canine; cell lines; cell culture; marker expression
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Hammer, S.C.; Becker, A.; Rateitschak, K.; Mohr, A.; Lüder Ripoli, F.; Hennecke, S.; Junginger, J.; Hewicker-Trautwein, M.; Brenig, B.; Ngezahayo, A.; Nolte, I.; Murua Escobar, H. Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines. Int. J. Mol. Sci. 2016, 17, 1655.

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