Department of Small Animal Clinical Sciences, University of Florida College of Veterinary Medicine, 2015 SW 16th Ave, Gainesville, FL 32608, United States.
Department of Small Animal Clinical Sciences, University of Florida College of Veterinary Medicine, 2015 SW 16th Ave, Gainesville, FL 32608, United States.
Vet Immunol Immunopathol. 2023 May;259:110594. doi: 10.1016/j.vetimm.2023.110594. Epub 2023 Apr 10.
GD2 and GD3 are disialoganglioside oncofetal antigens important in oncogenesis. GD2 synthase (GD2S) and GD3 synthase (GD3S) are needed for GD2 and GD3 production. The objectives of this study are to validate the use of RNA in situ hybridization (RNAscope®) in the detection of GD2S and GD3S in canine histiocytic sarcoma (HS) in vitro and optimize this technique in canine formalin-fixed paraffin-embedded (FFPE) tissues. A secondary objective is to evaluate the prognostic significance of GD2S and GD3S on survival. Quantitative RT-PCR compared GD2S and GD3S mRNA expression between three HS cell lines followed by RNAscope® in fixed cell pellets from the DH82 cell line and FFPE tissues. Variables prognostic for survival were determined with Cox proportional hazard model. RNAscope® was validated for detection of GD2S and GD3S and optimized in FFPE tissues. GD2S and GD3S mRNA expression was variable between cell lines. GD2S and GD3S mRNA expression was detected and measured in all tumor tissues; there was no association with prognosis. GD2S and GD3S are expressed in canine HS and successfully detected using the high throughput technique of RNAscope® in FFPE samples. This study provides the foundation for future prospective research of GD2S and GD3S utilizing RNAscope®.
GD2 和 GD3 是二唾液酸神经节苷脂肿瘤相关的胚胎抗原，在肿瘤发生中很重要。GD2 合酶 (GD2S) 和 GD3 合酶 (GD3S) 是 GD2 和 GD3 产生所必需的。本研究的目的是验证 RNA 原位杂交 (RNAscope®) 在体外检测犬组织细胞肉瘤 (HS) 中 GD2S 和 GD3S 的应用，并优化该技术在犬福尔马林固定石蜡包埋 (FFPE) 组织中的应用。次要目标是评估 GD2S 和 GD3S 对生存的预后意义。定量 RT-PCR 比较了三种 HS 细胞系中的 GD2S 和 GD3S mRNA 表达，然后用 RNAscope®检测 DH82 细胞系的固定细胞沉淀和 FFPE 组织。采用 Cox 比例风险模型确定与生存相关的预后变量。验证了 RNAscope®检测 GD2S 和 GD3S 的方法，并优化了 FFPE 组织中的检测方法。GD2S 和 GD3S 在细胞系之间的 mRNA 表达是可变的。GD2S 和 GD3S mRNA 表达在所有肿瘤组织中均被检测到；与预后无关联。GD2S 和 GD3S 在犬 HS 中表达，并成功地利用 FFPE 样本中的高通量 RNAscope®技术进行检测。本研究为未来利用 RNAscope®研究 GD2S 和 GD3S 的前瞻性研究提供了基础。
