Nelson Peter T, Baldwin Don A, Kloosterman Wigard P, Kauppinen Sakari, Plasterk Ronald H A, Mourelatos Zissimos
Division of Neuropathology and Department of Pathology & Laboratory Medicine, University of Pennsylvania School of Medicine, 605A Stellar-Chance Bldg., 421 Curie Blvd., Philadelphia, PA 19104, USA.
RNA. 2006 Feb;12(2):187-91. doi: 10.1261/rna.2258506. Epub 2005 Dec 22.
microRNAs (miRNAs) are small (approximately 22 nucleotide) regulatory RNAs which play fundamental roles in many biological processes. Recent studies have shown that the expression of many miRNAs is altered in various human tumors and some miRNAs may function as oncogenes or tumor suppressor genes. However, with the exception of glioblastoma multiforme, the expression of miRNAs in brain tumors is unknown. Furthermore, methods to profile miRNAs from formalin-fixed, paraffin-embedded (FFPE) archival tissues or to study their cellular and subcellular localization in FFPE tissues have been lacking. Here we report the coordinated miRNA expression analysis from the tissue level to the subcellular level, using the RAKE (RNA-primed, array-based, Klenow Enzyme) miRNA microarray platform in conjunction with Locked Nucleic Acid (LNA)-based in situ hybridization (LNA-ISH) on archival FFPE human brains and oligodendroglial tumors. The ability to profile miRNAs from archival tissues at the tissue level, by RAKE microarrays, and at the cellular level by LNA-ISH, will accelerate studies of miRNAs in human diseases.
微小RNA（miRNA）是小的（约22个核苷酸）调节性RNA，在许多生物学过程中发挥着重要作用。最近的研究表明，许多miRNA在各种人类肿瘤中的表达发生了改变，一些miRNA可能作为癌基因或肿瘤抑制基因发挥作用。然而，除多形性胶质母细胞瘤外，脑肿瘤中miRNA的表达情况尚不清楚。此外，一直缺乏从福尔马林固定、石蜡包埋（FFPE）存档组织中分析miRNA谱或研究其在FFPE组织中的细胞及亚细胞定位的方法。在此，我们报告了利用RAKE（RNA引发、基于微阵列、klenow酶）miRNA微阵列平台结合基于锁核酸（LNA）的原位杂交（LNA-ISH）技术，对存档的FFPE人脑和少突胶质细胞瘤进行从组织水平到亚细胞水平的协调miRNA表达分析。通过RAKE微阵列在组织水平以及通过LNA-ISH在细胞水平对存档组织中的miRNA进行分析的能力，将加速对人类疾病中miRNA的研究。
