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Abstract
RNA is considered as an indicator of the dynamic genetic expression changes in a cell. RNAScope is a commercially available in situ hybridization assay for the detection of RNA in formalin-fixed paraffin-embedded tissue. In this work, we describe the use of RNAScope as a sensitive and specific method for the evaluation of c-KIT messenger RNA (mRNA) in canine mast cell tumor. We investigated the expression of c-KIT mRNA with RNAscope in 60 canine mast cell tumors (MCTs), comparing it with the histological grade and KIT immunohistochemical expression patterns. Our results showed an overall good expression of c-KIT mRNA in neoplastic cells if compared with control probes. We also observed a statistically significant correlation between histological grade and c-KIT mRNA expression. No correlations were found between KIT protein immunohistochemical distribution pattern and c-KIT mRNA expression or histological grade. Our results provide a reference basis to better understand c-KIT mRNA expression in canine MCTs and strongly encourage further studies that may provide useful information about its potential and significant role as a prognostic and predictive biological marker for canine MCTs clinical outcome.
Highlights
Canine mast cell tumor (MCT) is one of the most common neoplastic disease in dogs accounting for ∼20% of all canine skin tumors [1, 2]
These results confirm that tissue microarray (TMA) constructed from a retrospective formalin fixed paraffin-embedded (FFPE) tissue archive series are fit-for-purpose for the evaluation of c-KIT messenger RNA (mRNA) by RNAScope in canine MCTs; for several reasons, this finding represent a different-making option in biomarker development and discovery
We need to address an important critical issue encountered during our experiments in order to offer useful suggestion to optimize the reproducibility of this assay on FFPE canine MCTs
Summary
Canine mast cell tumor (MCT) is one of the most common neoplastic disease in dogs accounting for ∼20% of all canine skin tumors [1, 2]. Cutaneous MCTs can have a very variable biological behavior [1], and several studies have focused on the investigation of predictive factors for MCTs outcome. These factors include the location of the tumor [3, 4], surgical margins [5,6,7,8,9], mitotic activity [10, 11], nuclear morphometry [12, 13], and vascular density [14]. Immunohistochemical expression and protein localization of KIT in neoplastic cells is currently one of the most informative markers for prognostication of canine MCTs [1, 15,16,17]. It has been extensively described that the immunohistochemical localization of KIT protein in neoplastic canine mast cells may have three different patterns: perimembranous labeling (pattern I), focal or stippled cytoplasmic labeling along with loss of perimembranous labeling (pattern II), and diffuse cytoplasmic labeling
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