Changes in differential gene expression because of warm ischemia time of radical prostatectomy specimens
Changes in differential gene expression because of warm ischemia time of radical prostatectomy specimens
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DOI：
10.1016/s0002-9440(10)64451-3
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发表时间：
2002-11-01
影响因子：
6
通讯作者：
Rubin, MA
中科院分区：
文献类型：
--
作者：
Dash, A;Maine, IP;Rubin, MA
利用cDNA微阵列技术可以同时监测数千个基因的表达。这项技术正被用于了解人类疾病的复杂性。一个重要的技术关注点是由于组织缺血的影响而可能导致的基因表达改变。本研究评估了因组织处理时间而导致的差异基因表达的增加。为了评估因缺血时间导致的差异基因表达，前列腺样本被分为五个时间点（0、0.5、1、3和5小时）。每个时间点由12到15个前列腺组织小块（5立方毫米）均匀混合而成。这些组织在室温下保存，直到在指定的时间点将组织置于OCT中，在液氮中速冻，并在-80℃下储存直至提取RNA。每个时间点的RNA与来自同一前列腺的0时间点RNA的一份进行杂交。四个前列腺腺体用于平行研究。绘制M - A图以比较时间点样本杂交之间的变异性。使用微阵列统计分析软件来识别在1小时时间点相较于0小时时间点过表达且具有统计学意义的基因。微阵列分析显示只有一小部分基因（
The expression of thousands of genes can be monitored simultaneously using cDNA microarray technology. This technology is being used to understand the complexity of human disease. One significant technical concern regards potential alterations in gene expression because of the effect of tissue ischemia. This study evaluates the increase in the differential gene expression because of tissue processing time. To evaluate differential gene expression because of ischemia time, prostate samples were divided into five time points (0, 0.5, 1, 3, and 5 hours). Each time point consisted of a homogeneous mixture of 12 to 15 prostate tissue cubes (5 mm(3)). These tissues were maintained at room temperature until at the assigned time point the tissue was placed in OCT, flash frozen in liquid nitrogen, and stored at -80degreesC until RNA extraction. RNA from each time point was hybridized against an aliquot of 0 time point RNA from the same prostate. Four prostate glands were used in parallel studies. M-A plots were graphed to compare variability between time point sample hybridizations. Statistical Analysis of Microarray software was used to identify genes overexpressed at the I-hour time point versus the 0-hour time with statistically significance. Microarray analysis revealed only a small percentage of genes (