Fluorescence In Situ Hybridization for MicroRNA Detection in Archived Oral Cancer Tissues.
Fluorescence In Situ Hybridization for MicroRNA Detection in Archived Oral Cancer Tissues.
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DOI：
10.1155/2012/903581
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发表时间：
2012
影响因子：
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通讯作者：
Stack MS
中科院分区：
文献类型：
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作者：
Shi Z;Johnson JJ;Stack MS
被称为 microRNA (miRNA) 的非编码 RNA 是一大类小型单链调节 RNA，在人类疾病研究中引起了广泛的兴趣。为了便于描述 microRNA 在癌症病理学中的作用，我们试图探索检测福尔马林固定石蜡包埋 (FFPE) 组织中 microRNA 表达的可行性。使用 FFPE 材料，我们比较了检测 miR-146a 的荧光原位杂交 (FISH) 程序与 (a) 不同的合成探针：常规定制 DNA 寡核苷酸与锁核酸 (LNA) 掺入 DNA 寡核苷酸； (b) 探针的不同报告基因：生物素与地高辛 (DIG)； (c) 不同的可视化：传统与酪酰胺信号放大（TSA）系统； (d) 内源过氧化物酶的不同封闭剂。最后，我们对市售的口腔癌组织微阵列进行了 miR-146a FISH，其中包含 40 例口腔鳞状细胞癌 (OSCC) 和 10 例来自人类口腔的正常上皮细胞。提供了用于检测 miR-146a 的 FISH 方案示例。总之，我们已经建立了可靠的原位杂交程序来检测 FFPE 口腔癌组织中 microRNA 的表达。该方法是研究 microRNA 参与口腔癌病理学的重要工具，可能具有潜在的预后或诊断价值。
The noncoding RNA designated as microRNA (miRNA) is a large group of small single-stranded regulatory RNA and has generated wide-spread interest in human disease studies. To facilitate delineating the role of microRNAs in cancer pathology, we sought to explore the feasibility of detecting microRNA expression in formalin-fixed paraffin-embedded (FFPE) tissues. Using FFPE materials, we have compared fluorescent in situ hybridization (FISH) procedures to detect miR-146a with (a) different synthetic probes: regular custom DNA oligonucleotides versus locked nucleic acid (LNA) incorporated DNA oligonucleotides; (b) different reporters for the probes: biotin versus digoxigenin (DIG); (c) different visualization: traditional versus tyramide signal amplification (TSA) system; (d) different blocking reagents for endogenous peroxidase. Finally, we performed miR-146a FISH on a commercially available oral cancer tissue microarray, which contains 40 cases of oral squamous cell carcinoma (OSCC) and 10 cases of normal epithelia from the human oral cavity. A sample FISH protocol for detecting miR-146a is provided. In summary, we have established reliable in situ hybridization procedures for detecting the expression of microRNA in FFPE oral cancer tissues. This method is an important tool for studies on the involvement of microRNA in oral cancer pathology and may have potential prognostic or diagnostic value.