Equine penile squamous cell carcinoma: expression of biomarker proteins and EcPV2

Abstract

equine penile squamous cell carcinoma (epScc) is a relatively common cutaneous neoplasm with a poor prognosis. in this study, we aimed to determine the protein expression and colocalisation of FRA1, c-Myc, Cyclin D1, and MMP7 in normal (NT), tumour (T), hyperplastic epidermis and/or squamous papilloma (Hyp/Pap), poorly-differentiated (PDSCC), or well-differentiated (WDSCC) EpSCC using a tissue array approach. further objectives were to correlate protein expression to (i) levels of inflammation, using a convolutional neural network (ii) equine papillomavirus 2 (EcPV2) infection, detected using PCR amplification. We found an increase in expression of FRA1 in EpSCC compared to NT samples. c-Myc expression was higher in Hyp/Pap and WDSCC but not PDSCC whereas MMP7 was reduced in WDSCC compared with NT. There was a significant increase in the global intersection coefficient (GIC) of FRA1 with MMP7, c-Myc, and Cyclin D1 in EpSCC. Conversely, GIC for MMP7 with c-Myc was reduced in EpSCC tissue. Inflammation was positively associated with EcPV2 infection in both NT and EpSCC but not Hyp/Pap. Changes in protein expression could be correlated with EcPV2 for Cyclin D1 and c-Myc. Our results evaluate novel biomarkers of EpSCC and a putative correlation between the expression of biomarkers, EcPV2 infection and inflammation. Equine penile squamous cell carcinoma (EpSCC) is a cutaneous neoplasm with a poor prognosis that often results in euthanasia due to late presentation, treatment difficulties and deterioration. EpSCC are often seen with precancerous pink to yellow plaques and genital papillomas. The lesion is seen mostly at the end of the second and beginning of the third decade of life 1. The term penile intraepithelial neoplasia (PIN) used in humans may also be applied to these lesions. After sarcoids, squamous cell carcinomas are considered the most common equine neoplasm 1-3. Around one tenth of all equine neoplasms are diagnosed in the penis, vulva and ocular adnexa 4,5 of which EpSCC is the most common. Incidence rates of EpSCC, reported more in ponies compared to horses 6 , vary and no specific breed predilection has been ascertained 6. The recorded incidence rates for EpSCCs are between 50-80% of all external genital neoplasms, however one report recorded that EpSCC made up around a fifth of all diagnosed equine cancers in a single UK laboratory over a 29-year period, with the incidence of cutaneous equine tumours also varying by region 6. The possible causes of EpSCC are suggested to be smegma accumulation, ultraviolet light overexposure, chronic irritation and balanoposthitis 7. Chronic inflammation is a known risk factor for cancer development 8. It is also thought that a majority of solid tumours are infiltrated with immune and inflammatory cells 9. The link between human papilloma virus (HPV), cervical cancer 10 and chronic inflammation is known 8. There is evidence to suggest that equine cancers may be initiated, in part, by papillomavirus infection analogous to human cervical and penile cancer 11. These sugge...

Comparative analysis of Wnt signaling-related proteins in normal, benign, malignant and metastasised human liver tumors

“…We have previously studied the quantitative expression of Wnt-related proteins in several cancers based on multi-labeled qIHC approach using TAs. (18,19) In this study, we used a Table S5). However, we observed significantly reduced expression of Cyclin D1 only in iCC, pCC, HB and PAC-LM.…”

“…Immunostaining was performed using BOND™ automated staining system (Leica Biosystems, Milton Keynes, UK) and BOND Polymer Refine Detection kit (DS9800, Leica Biosystems, Newcastle, UK) according to manufacturer’s recommendations, as described elsewhere. (18, 19) The list of the antibodies used and the optimisation of antibody staining process are given in Supporting Table S4 and Fig. S2.…”

“…(9,10) The heterogeneity of liver cancer types is an impediment for diagnosis and prognosis and there is a need to identify specific biomarkers for liver cancer subtypes in order to facilitate diagnosis and prognosis. Here we investigated the expression of four Wnt transcriptional targets, Cyclin D1, c-Myc, Fra-1 and Pygo-1, across different types of human liver tumors using an unbiased, qIHC approach (18,19) in a large number of tissue samples on tissue arrays (TAs). This is the first report investigating a number of downstream Wnt signaling transcriptional targets in a comparative analysis of liver tumors using qIHC and may provide insights into the mechanisms involved in specific liver tumors and disease outcome.…”

“…(13) An important endpoint of Wnt-mediated β-catenin stabilisation and translocation into the nucleus is the control of gene transcription via the nuclear TCF/LEF/β-catenin/Legless/Pygo complex; (14) a number of these genes are also transcription factors and include, c-Myc, (15) Cyclin D1, (16) and Fra-1. (17) Here we investigated the expression of four Wnt transcriptional targets, Cyclin D1, c-Myc, Fra-1 and Pygo-1, across different types of human liver tumors using an unbiased, qIHC approach (18, 19) in a large number of tissue samples on tissue arrays (TAs). This is the first report investigating a number of downstream Wnt signaling transcriptional targets in a comparative analysis of liver tumors using qIHC and may provide insights into the mechanisms involved in specific liver tumors and disease outcome.…”

Experimental Models for Studying HPV-Positive and HPV-Negative Penile Cancer: New Tools for An Old Disease

“…Another interesting model for studying penile cancer induced by papillomaviruses may be found in horses. These animals develop penile papillomas, intraepithelial neoplastic lesions and squamous cell carcinomas which are associated with infection by equine papillomavirus type 2 (EcPV2) [ 79 , 98 , 99 , 100 ]. Importantly, some equine penile squamous cell carcinomas are able to metastasize to regional lymph nodes, something that has not yet been observed when studying the newly available mouse models [ 79 ].…”