Joint Graduate School of Veterinary Medicine, Kagoshima University, Korimoto, Kagoshima, Japan.
Veterinary Teaching Hospital, Joint Faculty of Veterinary Medicine, Kagoshima University, Korimoto, Kagoshima, Kagoshima, Japan.
Vet Q. 2024 Dec;44(1):1-8. doi: 10.1080/01652176.2023.2300943. Epub 2024 Jan 30.
Hypoxia may promote tumor progression, and hypoxically altered noncoding RNA (ncRNA) expression may play a role in metastasis. Canine oral melanoma (COM) frequently metastasizes, and ncRNA expression under hypoxia may be clinically significant. We aimed to elucidate ncRNA fragments whose expression is altered by hypoxia in COM-derived primary KMeC and metastatic LMeC cell lines using next-generation sequencing to validate these results in qRT-PCR, and then compare expression between metastatic and non-metastatic COM. The NGS analysis and subsequent qRT-PCR validation were performed using hypoxic and normoxic KMeC and LMeC cells, and clinical samples [tumor tissue, plasma, and plasma-derived extracellular vesicles] obtained from dogs with metastatic or non-metastatic melanoma were analyzed with qRT-PCR. Y RNA was significantly decreased in metastatic LMeC cells versus primary KMeC cells in hypoxic and normoxic conditions. The expression of Y RNA was decreased in dogs with metastatic melanoma versus those with non-metastatic melanoma for all clinical sample types, reflecting the pattern found with hypoxia. Receiver operating characteristic analysis demonstrated that Y RNA level is a promising biomarker for discriminating metastatic from non-metastatic melanoma in plasma [area under the curve (AUC) = 0.993, < 0.0001] and plasma-derived extracellular vesicles (AUC = 0.981, = 0.0002). Overall, Y RNA may be more resistant to hypoxic stress in the metastatic than the non-metastatic state for COM. However, further investigation is required to elucidate the biological functions of Y RNA under hypoxic conditions.
缺氧可能促进肿瘤进展，而缺氧改变的非编码 RNA（ncRNA）表达可能在转移中发挥作用。犬口腔黑色素瘤（COM）经常转移，ncRNA 在缺氧下的表达可能具有临床意义。我们旨在阐明通过下一代测序在 COM 衍生的原发性 KMeC 和转移性 LMeC 细胞系中由缺氧改变的 ncRNA 片段的表达，并用 qRT-PCR 验证这些结果，然后比较转移和非转移 COM 之间的表达。使用缺氧和常氧 KMeC 和 LMeC 细胞进行 NGS 分析和随后的 qRT-PCR 验证，并使用 qRT-PCR 分析来自患有转移性或非转移性黑色素瘤的狗的肿瘤组织、血浆和血浆衍生的细胞外囊泡等临床样本。在缺氧和常氧条件下，转移性 LMeC 细胞中的 Y RNA 明显低于原发性 KMeC 细胞。与非转移性黑色素瘤相比，患有转移性黑色素瘤的狗的 Y RNA 表达在所有临床样本类型中均降低，反映了缺氧时的模式。接受者操作特征分析表明，Y RNA 水平是区分血浆中转移性和非转移性黑色素瘤的有前途的生物标志物 [曲线下面积（AUC）= 0.993， < 0.0001] 和血浆衍生的细胞外囊泡（AUC = 0.981， = 0.0002）。总体而言，与非转移性状态相比，Y RNA 在转移性 COM 中可能对缺氧应激更具抵抗力。然而，需要进一步研究来阐明 Y RNA 在缺氧条件下的生物学功能。
