Department of Laboratory Medicine and Children's and Women's Health, Norwegian University of Science and Technology, Trondheim, Norway.
Scand J Clin Lab Invest. 2010 Feb;70(1):45-53. doi: 10.3109/00365510903540815.
High-throughput technologies such as microarray have enhanced the discovery of new biomarkers in prostate cancer. However, the reliability of transcriptome analyses is limited by the RNA quality.
Identification of variables influencing the RNA quality in radical prostatectomy specimens.
RNA was extracted using an automatic extraction method for 354 samples from 38 fresh frozen prostate slices, and by manual extraction for 28 samples from 5 slices. RNA quality was measured using the RIN method (RNA Integrity Number). Evaluation of tissue composition was performed by light-microscopy for each sample. Age, total operative time, estimated blood loss, prostate volume, prostate specific antigen (s-PSA) and postoperative Gleason score were registered. The independent variables were correlated to the RIN score in a multiple linear regression model, taking p < 0.05 as the significance limit.
The amount of blood loss during prostatectomy and the amount of stroma in the tissue sample both correlated negatively with the RIN score (p = 0.03 and 0.02). Automatically extracted samples which were exposed to heat according to the RNA extraction protocol, had lower mean RNA quality (5.5, 1.46 SD) than manually extracted samples, not exposed to heat (8.7, 0.86 SD), suggesting degradation by temperature sensitive RNases, mainly residing in the stroma.
The highest RNA quality isolated by an automatic method from fresh frozen prostate tissue is obtained from patients with low peroperative blood loss and from samples with a low stromal fraction.
高通量技术如微阵列增强了前列腺癌新生物标志物的发现。然而，转录组分析的可靠性受到 RNA 质量的限制。
鉴定影响根治性前列腺切除术标本中 RNA 质量的变量。
使用自动提取方法从 38 个新鲜冷冻前列腺切片中提取了 354 个样本的 RNA，从 5 个切片中提取了 28 个样本的 RNA 进行手动提取。使用 RIN 法（RNA 完整性编号）测量 RNA 质量。对每个样本进行组织成分的光镜评估。记录年龄、总手术时间、估计失血量、前列腺体积、前列腺特异性抗原（s-PSA）和术后 Gleason 评分。将独立变量与 RIN 评分进行多元线性回归模型分析，以 p<0.05 为显著性界限。
前列腺切除术期间的失血量和组织样本中的基质量与 RIN 评分呈负相关（p=0.03 和 0.02）。根据 RNA 提取方案自动提取的样本，与未受热的手动提取样本相比，平均 RNA 质量较低（5.5，1.46 SD），表明温度敏感的 RNase 降解，主要存在于基质中。
从新鲜冷冻的前列腺组织中通过自动方法分离得到的最高 RNA 质量，来自于术中失血量低的患者和基质比例低的样本。
