Tsumoto Shohei, Tamura Kyoichi, Nakazawa Yuta, Fujita Michio, Ohkusu-Tsukada Kozo
Laboratory of Veterinary Pathology, School of Veterinary Medicine, Nippon Veterinary and Life-Science University (NVLU), Tokyo, Japan.
Laboratory of Veterinary Clinical Pathology, School of Veterinary Medicine, Nippon Veterinary and Life-Science University (NVLU), Tokyo, Japan.
Front Vet Sci. 2025 Mar 27;12:1552415. doi: 10.3389/fvets.2025.1552415. eCollection 2025.
Animal practice requires both convenience for the owner and risk management for the animal's health. Deterioration due to cancer may associate with poor prognosis under general anesthesia, which need to partial excision for pathological diagnosis. This study aimed to establish rapidly detecting the expression of survivin antigens for cancer vaccines or molecular targeted therapies via flow cytometry (FCM) using the intracellular staining method in tumor samples obtained via needle biopsy without anesthesia. Therefore, survivin expression patterns in each cell lines of canine melanomas, a murine mast cell tumor, a murine colon carcinoma, and a murine melanoma was analyzed by FCM and immunofluorescence microscopy, and compared with immunohistochemical analysis and western blot method. Interestingly, FCM results of the bimodal expression pattern of survivin were suggested to reflect the high fluorescence intensity of its nuclear-cytosol localization and the weak fluorescence intensity of its cytosol alone localization. In a case of canine cancer disease, it was confirmed that survivin expression patterns can be detected via FCM using needle biopsy samples in actual clinical settings. In this study, a novel method via FCM was proposed to quickly determine also survivin localization not only whether the survivin is expressed in cancer cells. The application of cancer vaccine or chemical therapy via this technology can be expected to contribute to improved animal care due to the "one-day first program," which has been proposed in convenience for owners.
动物医疗实践既需要方便主人,又需要对动物健康进行风险管理。癌症导致的病情恶化可能与全身麻醉下的不良预后相关,这需要进行部分切除以进行病理诊断。本研究旨在通过流式细胞术(FCM),采用细胞内染色方法,在未经麻醉的针吸活检获得的肿瘤样本中,快速检测癌症疫苗或分子靶向治疗中生存素抗原的表达。因此,通过FCM和免疫荧光显微镜分析了犬黑色素瘤、小鼠肥大细胞瘤、小鼠结肠癌和小鼠黑色素瘤各细胞系中的生存素表达模式,并与免疫组织化学分析和蛋白质印迹法进行了比较。有趣的是,生存素双峰表达模式的FCM结果表明,其反映了核-胞质定位的高荧光强度和单独胞质定位的弱荧光强度。在一例犬类癌症病例中,证实了在实际临床环境中,使用针吸活检样本通过FCM可以检测到生存素表达模式。在本研究中,提出了一种通过FCM的新方法,不仅可以快速确定癌细胞中是否表达生存素,还可以确定其定位。由于为方便主人而提出的“一日优先计划”,预计通过该技术应用癌症疫苗或化学疗法有助于改善动物护理。