NGS-identified miRNAs in Canine Mammary Gland Tumors Show Unexpected Expression Alterations in qPCR Analysis
NGS-identified miRNAs in Canine Mammary Gland Tumors Show Unexpected Expression Alterations in qPCR Analysis
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DOI：
10.21873/invivo.12873
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发表时间：
2022-07-01
期刊：
影响因子：
2.3
通讯作者：
Miura, Naoki
中科院分区：
文献类型：
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作者：
Chen, Hui-Wen;Lai, Yu-Chang;Miura, Naoki
背景/目的：犬乳腺肿瘤（MGT）作为人类乳腺癌的潜在模型，有一个明确的组织学分类系统。微小RNA（miRNA）表达是MGT和人乳腺癌的分子特征的关键部分，尽管单独的特征还不能为明确的诊断提供足够的基础。在这项研究中，我们研究了miRNA表达模式和组织学分类之间的关联。材料与方法：取健康犬（n=7）和患乳腺癌犬（n=80）的乳腺组织。从建立的MGT细胞系获得另外的样品（n=5）。我们靶向转移性肿瘤组织与非转移性和正常组织中差异表达的miRNA。使用小RNA下一代测序（NGS）和随后的qPCR分析样品的子集。结果：从NGS分析中选择了6个差异表达的miRNAs，并提交用于大规模qPCR。大规模qPCR分析揭示了miRNA表达的更大变化。基于79个样本的大规模分析揭示了基于所选miRNA的分层聚类，其与组织病理学亚型分类并不显著匹配。结论：成功地研究了犬MGT中大规模miRNA的表达模式，并提供了完整的miRNA表达。选择的miRNA表明，在miRNA水平上，犬MGT的分子特征和组织学分类之间没有直接的映射。
Background/Aim: Canine mammary gland tumors (MGTs), as a potential model of human breast cancer, have a well-defined histological classification system. MicroRNA (miRNA) expression is a key part of the molecular signatures of both MGTs and human breast cancer, although the signatures alone do not yet provide a sufficient basis for definitive diagnosis. In this study, we investigated the association between miRNA expression patterns and histological classification. Materials and Methods: Mammary gland tissue was collected from healthy dogs (n=7) and dog patients (n=80). Further samples (n=5) were obtained from established MGT cell lines. We targeted miRNAs differentially expressed in metastatic tumor tissue versus non-metastatic and normal tissue. A subset of samples was analyzed using small RNA next generation sequencing (NGS) with subsequent qPCR. Results: Six differentially expressed miRNAs were selected from the NGS analysis and submitted for large-scale qPCR. The large-scale qPCR analysis revealed greater alternations in miRNA expression. Large-scale analysis, based on 79 samples, revealed a hierarchical clustering based on selected miRNAs that did not strikingly match the histopathological subtype classification. Conclusion: We successfully investigated the large-scale miRNA expression pattern in canine MGT and provided the whole miRNA expression. The selected miRNA demonstrated that there is no straightforward mapping between molecular signatures and histological classification of canine MGTs at the miRNA level.