Department of Biobank, Renji Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200127, China.
Anal Biochem. 2012 Apr 15;423(2):229-35. doi: 10.1016/j.ab.2012.02.003. Epub 2012 Feb 13.
Surgically removed samples of high quality provide more accurate and reliable results in downstream molecular assays. Some factors, including the type of anesthesia, surgical manipulation, transport time and mode, preservation method, storage length, and number of freeze-thaw cycles, can affect biosample quality and the subsequent gene expression analysis. Warm ischemia resulting from these factors has a substantial effect on biosample quality and is the focus of this mini review. We classified the effects of warm ischemia on gene expression as (i) warm ischemia-induced metabolic activity (WIMA) in living cells and (ii) warm ischemia-induced RNA degradation (WIRD). The differential effects of WIMA and WIRD on gene expression analysis appear to depend on the period after surgical removal. WIMA predominantly affects gene expression during the early stage after surgery, whereas WIRD has a more significant effect after tissue thawing. By a literature review, we also found that RNA isolated from surgically removed biopsies is stable, and high-quality RNA can be obtained for most nonfixed human tissue maintained at room temperature during the early period after surgery. Understanding these characteristics of gene expression variation should help biomedical researchers to avoid misleading gene expression results.
高质量的手术切除样本可为下游分子检测提供更准确和可靠的结果。一些因素，包括麻醉类型、手术操作、运输时间和方式、保存方法、储存时间以及冻融循环次数，都会影响生物样本的质量和随后的基因表达分析。这些因素导致的热缺血对生物样本质量有重大影响，是本综述的重点。我们将热缺血对基因表达的影响分为（i）活细胞中的热缺血诱导代谢活性（WIMA）和（ii）热缺血诱导的 RNA 降解（WIRD）。WIMA 和 WIRD 对基因表达分析的不同影响似乎取决于手术后的时间。WIMA 主要影响手术后早期的基因表达，而 WIRD 在组织解冻后影响更大。通过文献回顾，我们还发现，从手术切除的活检中分离的 RNA 是稳定的，在手术后的早期，大多数非固定的人类组织在室温下仍可获得高质量的 RNA。了解这些基因表达变化的特征应有助于生物医学研究人员避免误导性的基因表达结果。
