Novel Rank-Based Statistical Methods Reveal MicroRNAs with Differential Expression in Multiple Cancer Types. Homo sapiens

The data collected in this study consisted of 32 paired samples of cancerous and normal origins from 14 different patients and 8 different cancer types. Each pair of samples was taken from different parts of the same tissue in the same patient - one collected and extracted from the tumor and one collected and extracted from adjacent normal tissue. One pair from each of the breast, lymphoma and prostate tissues, 2 pairs from liver and lung tissues and 3 pairs from colon, ovary and testes tissues. 2 of the ovary and testes pairs were technical replicates. We use commercial adjacent tissue tumor-normal total RNA samples purchased from Ambion/ABI. The samples were hybridized to Agilent's miRNA microarray version 2.0. Keywords: miRNA Overall design: For each microarray analysis, 100ng total RNAs were labeled with Cy3 per Agilent miRNA Micorarray Systems Protocol v1.5. The labeled RNA samples were hybridized onto Agilent miRNA microarray containing 799 miRNAs(Agilent Human miRNA Microarray kit, V2) for 21 hours at 55C. The arrays were then washed at room temperature and scanned to produce the hybridization signals (Agilent miRNA Micorarray Systems Protocol v1.5). The arrays were scanned with extended dynamic range at 5 and 100% PMT using the Agilent scanner (model G2565AA Technical replicate: Sample 7, Sample 29 Technical replicate: Sample 8, Sample 30 Technical replicate: Sample 19, Sample 31 Technical replicate: Sample 20, Sample 32

本研究采集的数据集包含14名不同患者、8种不同癌症类型的32对癌组织与正常组织配对样本。每对样本均取自同一名患者同一组织的两个不同部位：一份提取自肿瘤组织，另一份提取自相邻正常组织。其中乳腺、淋巴瘤和前列腺组织各对应1对样本，肝、肺组织各对应2对样本，结肠、卵巢和睾丸组织各对应3对样本。卵巢与睾丸组织的配对样本中，有2对为技术重复样本。本研究使用了从Ambion/ABI公司采购的商品化相邻组织来源的肿瘤-正常配对总RNA样本。

样本按照安捷伦（Agilent）微小RNA（microRNA, miRNA）微阵列系统v1.5方案，使用Cy3对100ng总RNA进行荧光标记。标记完成的RNA样本与搭载799个miRNA的安捷伦人miRNA微阵列试剂盒V2在55℃下杂交21小时。随后在室温条件下洗涤芯片，并通过安捷伦扫描仪（型号G2565AA）扫描以获取杂交信号，扫描采用扩展动态范围，设置光电倍增管（Photomultiplier Tube, PMT）增益为5%和100%，所有操作均遵循安捷伦微小RNA（microRNA, miRNA）微阵列系统v1.5方案。

关键词：微小RNA（microRNA, miRNA）
实验设计概述：针对每一次微阵列分析，均按照安捷伦微小RNA（microRNA, miRNA）微阵列系统v1.5方案，使用Cy3标记100ng总RNA。标记后的RNA样本与搭载799个miRNA的安捷伦人miRNA微阵列试剂盒V2在55℃下杂交21小时。随后在室温下洗涤芯片并扫描以获取杂交信号，操作遵循安捷伦微小RNA（microRNA, miRNA）微阵列系统v1.5方案。扫描时采用扩展动态范围，使用安捷伦扫描仪（型号G2565AA），设置PMT增益为5%和100%。

技术重复对应关系如下：样本7与样本29、样本8与样本30、样本19与样本31、样本20与样本32