The HOPE-technique permits Northern blot and microarray analyses in paraffin-embedded tissues
The HOPE-technique permits Northern blot and microarray analyses in paraffin-embedded tissues
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DOI：
10.1016/j.prp.2004.04.008
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发表时间：
2004-01-01
影响因子：
2.8
通讯作者：
Bullerdiek, J
中科院分区：
文献类型：
--
作者：
Goldmann, T;Flohr, AM;Bullerdiek, J
在用于常规组织学检查之后，对组织样本的需求不断增加。也适用于分子分析。至于福尔马林固定的石蜡包埋组织，后者的应用非常有限。HOPE技术包括一种新的保护液，含有有机缓冲液，丙酮是唯一的脱水剂，纯石蜡的熔化温度为52-54℃，允许所有的病理常规检查。与FFPE组织相比，HOPE技术允许应用分子方法，如高分子DNA和RNA分离，可用于聚合酶链式反应和逆转录聚合酶链式反应(RT-PCR)。在这项研究中，我们调查了从HOPE固定的组织样本中提取的RNA是否适用于Northern印迹和微阵列分析。用两个不同组织学分级的HOPE固定的乳腺癌标本的RNA进行阵列实验。原来，从HOPE固定的组织中提取的RNA质量很高，可以成功地用于阵列实验。此外，通过检测GAPDH和高迁移率族蛋白基因B1(HMGB1)特异的转录本，我们能够证明来自HOPE固定组织的RNA也可以用于Northern印迹杂交。(C)2004年爱思唯尔股份有限公司。版权所有。
There is an increasing demand for tissue samples that, after having been used for conventional histologic examination. are also suited for molecular analyses. As to formalin-fixed, paraffin embedded (FFPE) tissue, the latter applications are very limited. The HOPE (Hepes-Glutamic acid buffer mediated Organic solvent Protection Effect) technique comprises a new protection-solution with an organic buffer, with acetone as the only dehydrating agent, and pure paraffin of 52-54degreesC melting temperature, allowing for all pathologic routine investigations. In contrast to FFPE tissue, the HOPE-technique allows for the application of molecular methods, such as high molecular DNA and RNA isolation, which can be used for PCR and reverse transcription PCR (RT-PCR). In this study, we investigated whether RNA from HOPE-fixed tissue samples is suitable for Northern blot and microarray analyses. RNAs of two HOPE-fixed breast cancer specimens of different histologic grade were used to carry out an array experiment. It turned out that RNA from HOPE-fixed tissue is of high quality and can be successfully used for array experiments. In addition, by detecting GAPDH and high mobility group protein gene B1 (HMGB1)-specific transcripts, we were able to demonstrate that RNA from HOPE-fixed tissue can also be used for Northern blot hybridization. (C) 2004 Elsevier GmbH. All rights reserved.