title: Quantitative Expression and Co-Localization of Wnt Signalling Related Proteins in Feline Squamous Cell Carcinoma.
abstract: Feline oral squamous cell carcinoma (FOSCC) is an aggressive neoplasm in cats. Little is known about the possible molecular mechanisms that may be involved in the initiation, maintenance and progression of FOSCC. Wnt signalling is critical in development and disease, including many mammalian cancers. In this study, we have investigated the expression of Wnt signalling related proteins using quantitative immunohistochemical techniques on tissue arrays. We constructed tissue arrays with 58 individual replicate tissue samples. We tested for the expression of four key Wnt/ß-catenin transcription targets, namely Cyclin D1 (CCND1 or CD1), FRA1, c-Myc and MMP7. All antibodies showed cross reactivity in feline tissue except MMP7. Quantitative immunohistochemical analysis of single proteins (expressed as area fraction / amount of tissue for normal vs tumor, mean ± SE) showed that the expression of CD1 (3.9 ± 0.5 vs 12.2 ± 0.9), FRA1 (5.5 ± 0.6 vs 16.8 ± 1.1) and c-Myc (5.4 ± 0.5 vs 12.5 ± 0.9) was increased in FOSCC tissue by 2.3 to 3 fold compared to normal controls (p<0.0001). By using a multilabel, quantitative fluorophore technique we further investigated if the co-localization of these proteins (all transcription factors) with each other and in the nucleus (stained with 4',6-diamidino-2-phenylindole, DAPI) was altered in FOSCC compared to normal tissue. The global intersection coefficients, a measure of the proximity of two fluorophore labeled entities, showed that there was a significant change (p < 0.01) in the co-localization for all permutations (e.g. CD1/FRA1 etc), except for the nuclear localization of CD1. Our results show that putative targets of Wnt signalling transcription are up-regulated in FOSCC with alterations in the co-localization of these proteins and could serve as a useful marker for the disease.
authors: Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom.
authors: Antonio
authors: A
authors: Giuliano
authors: Department of Clinical Sciences and Services, The Royal Veterinary College, London, United Kingdom.|Division of Surgery, University College London, London, United Kingdom.
authors: Rebecca
authors: R
authors: Swift
authors: Prostate Cancer Research Center at the Centre for Stem Cells and Regenerative Medicine, King's College London, London, United Kingdom.
authors: Callum
authors: C
authors: Arthurs
authors: Department of Clinical Sciences and Services, The Royal Veterinary College, London, United Kingdom.|Division of Surgery, University College London, London, United Kingdom.
authors: Georgina
authors: G
authors: Marote
authors: Department of Veterinary Sciences, University of Pisa, Pisa, Italy.
authors: Francesca
authors: F
authors: Abramo
authors: IDEXX Laboratories Ltd., Grange House, West Yorkshire, United Kingdom.
authors: Jenny
authors: J
authors: McKay
authors: Dundee Imaging Facility, College of Life Sciences, University of Dundee, Dundee, United Kingdom.
authors: Calum
authors: C
authors: Thomson
authors: Dundee Imaging Facility, College of Life Sciences, University of Dundee, Dundee, United Kingdom.
authors: Mariana
authors: M
authors: Beltran
authors: Queen's Medical Research Institute, University of Edinburgh, Edinburgh, United Kingdom.
authors: Michael
authors: M
authors: Millar
authors: Department of Pathology and Pathogen Biology, The Royal Veterinary College, London, United Kingdom.
authors: Simon
authors: S
authors: Priestnall
authors: Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom.
authors: Jane
authors: J
authors: Dobson
authors: Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom.
authors: Fernando
authors: F
authors: Costantino-Casas
authors: Prostate Cancer Research Center at the Centre for Stem Cells and Regenerative Medicine, King's College London, London, United Kingdom.
authors: Terry
authors: T
authors: Petrou
authors: Department of Comparative Biomedical Sciences, The Royal Veterinary College, London, United Kingdom.
authors: Imelda M
authors: IM
authors: McGonnell
authors: University of London, London, United Kingdom.
authors: Anthony J
authors: AJ
authors: Davies
authors: Independent Vetcare, Bath, United Kingdom.
authors: Malcolm
authors: M
authors: Weetman
authors: Department of Clinical Sciences and Services, The Royal Veterinary College, London, United Kingdom.
authors: Oliver A
authors: OA
authors: Garden
authors: Division of Surgery, University College London, London, United Kingdom.
authors: John R
authors: JR
authors: Masters
authors: Research Department of Cell and Developmental Biology, The Centre for Cell and Molecular Dynamics, Rockefeller Building, University College London, London, United Kingdom.
authors: Christopher
authors: C
authors: Thrasivoulou
authors: Prostate Cancer Research Center at the Centre for Stem Cells and Regenerative Medicine, King's College London, London, United Kingdom.|Division of Surgery, University College London, London, United Kingdom.
authors: Aamir
authors: A
authors: Ahmed
mesh_terms: D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000662:veterinary; D002371:Cat Diseases / Q000378:metabolism*; D002415:Cats; D019938:Cyclin D1 / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006863:Hydrogen-Ion Concentration; D020783:Matrix Metalloproteinase 7 / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009062:Mouth Neoplasms / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D012372:ROC Curve; D014157:Transcription Factors / Q000378:metabolism; D051153:Wnt Proteins / Q000378:metabolism; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000378:metabolism
chemical_list: D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D016271:Proto-Oncogene Proteins c-myc; D014157:Transcription Factors; D051153:Wnt Proteins; D051176:beta Catenin; D019938:Cyclin D1; D020783:Matrix Metalloproteinase 7