Genome-wide gene expression profiles of primary prostate cancer. Genome-wide gene expression profiles of primary prostate cancer

Prognostic biomarkers are useful to screen patients with clinically localized prostate cancer (PCa) who are at high risk of metastatic progression. The tumor transcriptome can be used to evaluate the aggressiveness of PCa and predict adverse patient outcomes. Genomewide gene expression levels were measured in primary tumor samples of 503 patients in a population‐based cohort. Overall design: Formalin‐fixed, paraffin‐embedded (FFPE) PCa tumor tissue blocks were obtained from radical prostatectomy samples and used to make H&E‐stained slides, which were reviewed by a PCa pathologist to confirm the presence of and the location of prostate adenocarcinoma. For each patient, two 1‐mm tumor tissue cores were taken from the areas enriched with ≥ 75% tumor cells from the dominant lesion. The RNeasy® FFPE Kit (Qiagen Inc., Valencia, CA, USA) was used to isolate the RNA from tissue cores, and the samples were quantified with RiboGreen, aliquoted (200 ng per patient) onto 96‐well plates, and shipped to Illumina for gene expression profiling. Tumor RNA samples from patients with various outcomes were randomly distributed across the plates and laboratory personnel were blinded to this information.

预后生物标志物（prognostic biomarkers）可用于筛查临床局限性前列腺癌（prostate cancer, PCa）中存在转移进展高风险的患者。肿瘤转录组（tumor transcriptome）可用于评估前列腺癌的侵袭性，并预测患者的不良临床结局。
本研究针对一项基于人群的队列，对503例患者的原发性肿瘤样本开展了全基因组基因表达水平检测。
实验整体设计如下：
福尔马林固定石蜡包埋（Formalin-fixed paraffin-embedded, FFPE）的前列腺癌肿瘤组织块取自根治性前列腺切除术样本，用于制作苏木精-伊红（Hematoxylin-Eosin, H&E）染色切片，随后由前列腺癌病理学家审阅切片以确认前列腺腺癌的存在及其病灶位置。针对每位患者，从其主要病灶中肿瘤细胞占比≥75%的富集区域采集两块直径1mm的肿瘤组织芯。使用RNeasy® FFPE试剂盒（Qiagen公司，美国加利福尼亚州巴伦西亚）从组织芯中提取RNA，通过RiboGreen法对样本进行定量，将每位患者200ng的样本分装至96孔板，随后送往Illumina公司进行基因表达谱分析。不同临床结局患者的肿瘤RNA样本被随机分配至各孔板中，实验人员对该分组信息采用盲法处理。