TY - JOUR
T1 - Quantitative Expression and Co-Localization of Wnt Signalling Related Proteins in Feline Squamous Cell Carcinoma
AU - Giuliano, Antonio
AU - Swift, Rebecca
AU - Arthurs, Callum
AU - Marote, Georgina
AU - Abramo, Francesca
AU - McKay, Jenny
AU - Thomson, Calum
AU - Beltran, Mariana
AU - Millar, Michael
AU - Priestnall, Simon
AU - Dobson, Jane
AU - Costantino-Casas, Fernando
AU - Petrou, Terry
AU - McGonnell, Imelda M
AU - Davies, Anthony J
AU - Weetman, Malcolm
AU - Garden, Oliver A
AU - Masters, John R
AU - Thrasivoulou, Christopher
AU - Ahmed, Aamir
PY - 2016/8/25
Y1 - 2016/8/25
N2 - 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.
AB - 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.
KW - Animals
KW - Carcinoma, Squamous Cell/metabolism
KW - Cat Diseases/metabolism
KW - Cats
KW - Cyclin D1/metabolism
KW - Gene Expression Regulation, Neoplastic
KW - Hydrogen-Ion Concentration
KW - Matrix Metalloproteinase 7/metabolism
KW - Microtubule-Associated Proteins/metabolism
KW - Mouth Neoplasms/metabolism
KW - Neoplasm Proteins/genetics
KW - Proto-Oncogene Proteins c-myc/metabolism
KW - ROC Curve
KW - Transcription Factors/metabolism
KW - Wnt Proteins/metabolism
KW - Wnt Signaling Pathway
KW - beta Catenin/metabolism
U2 - 10.1371/journal.pone.0161103
DO - 10.1371/journal.pone.0161103
M3 - Article (Academic Journal)
C2 - 27559731
SN - 1932-6203
VL - 11
JO - PLoS ONE
JF - PLoS ONE
IS - 8
M1 - e0161103
ER -