Abstract
Simple Summary5-aminolevulinic acid (5-ALA) is a natural amino acid that is metabolized in the mitochondria and used in the synthesis of heme. In human medicine, the fluorescence property of 5-ALA has been used for photodynamic tumor diagnosis. Nevertheless, research on 5-ALA in veterinary medicine is very limited. In this study, we aimed to evaluate the efficacy of 5-ALA to detect canine urothelial carcinoma and to clarify its diagnostic accuracy. The addition of 5-ALA to tumor cells from the urine of patients with urothelial carcinoma and canine urothelial carcinoma cell lines caused red fluorescence, and the amount of fluorescence was significantly higher than that in healthy dogs. Additionally, cases with high fluorescence intensity had more tumor invasion and metastasis. This study showed that 5-ALA can be used to detect canine urothelial carcinoma cells in urine with relatively high diagnostic accuracy.This study aimed to establish a method to detect canine urothelial carcinoma cells in urine using 5-aminolevulinic acid (5-ALA) and to evaluate its diagnostic accuracy. Urine samples were collected from 21 dogs diagnosed with urothelial carcinoma and three urothelial carcinoma cell lines were used. Urine samples obtained from seven healthy dogs were used as controls. Cells in the urine sediment, or urothelial carcinoma cell lines, were cultured with 5-ALA and then observed under a fluorescence microscope. Moreover, we examined the relationship between fluorescence intensity and the presence of metastasis as well as tumor invasion into the bladder wall in cases of urothelial carcinoma. Urine-derived cells from urothelial carcinoma and urothelial carcinoma cell lines showed clearer red fluorescence with the addition of 5-ALA compared to that exhibited by the cells from healthy dogs. The sensitivity and specificity of the diagnosis of urothelial carcinoma were 90% and 86%, respectively. Significant associations were found between fluorescence intensity and tumor metastasis and bladder wall invasion. This study showed that 5-ALA can be used to detect urothelial carcinoma cells in dogs with relatively high diagnostic accuracy. Further, the fluorescence intensity of tumor cells caused by 5-ALA correlated with the clinical condition of urothelial carcinoma cases, which suggested that 5-ALA could be used as a prognostic marker for canine urothelial carcinoma.
Highlights
Upon addition of 5-aminolevulinic acid (5-ALA), distinct red fluorescence was observed in the cytoplasm of three canine urothelial carcinoma cell lines: Sora, Love, and TCCUB
When the red fluorescence observed in these cases was quantified using a plate reader, the fluorescence ratio was significantly increased in urothelial carcinoma cases and urothelial carcinoma cell lines, compared to healthy dogs (Figure 2)
This study showed that urine-derived tumor cells could be detected with high sensitivity and specificity using 5-ALA in canine urothelial carcinoma
Summary
Known as transitional cell carcinoma, is a malignant tumor characterized by local invasion and metastasis to the lymph nodes and distant sites [1]. There is a need for effective medical treatment of urothelial carcinoma, as well as simple and accurate diagnostic methods for early detection of these tumors. The detection of abnormal epithelial cells in urine collected from patients by cytology can aid in diagnosis [2,3]. Cytology alone cannot diagnose canine urothelial carcinoma because normal urothelial cells can become morphologically abnormal, similar to tumor cells, due to prolonged contact with urine or inflammation [4]. The detection of BRAF mutations in urine-derived cells is widely used as a method to support the diagnosis of urothelial carcinoma in dogs. BRAF is a type of RAF protein belonging to the RAF/MEK/ERK mitogen-activated protein kinase (MAPK)
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