The Queensland Institute of Medical Research, Brisbane, Australia.
J Pathol. 2010 Aug;221(4):452-61. doi: 10.1002/path.2728.
Tissue sample acquisition is a limiting step in many studies. There are many thousands of formalin-fixed, paraffin-embedded archival blocks collected around the world, but in contrast relatively few fresh frozen samples in tumour banks. Once samples are fixed in formalin, the RNA is degraded and traditional methods for gene expression profiling are not suitable. In this study, we have evaluated the ability of the whole genome DASL (cDNA-mediated Annealing, Selection, extension, and Ligation) assay from Illumina to perform transcriptomic analysis of archived breast tumour tissue in formalin-fixed, paraffin-embedded (FFPE) blocks. We profiled 76 familial breast tumours from cases carrying a BRCA1, BRCA2 or ATM mutation, or from non-BRCA1/2 families. We found that replicate samples correlated well with each other (r(2) = 0.9-0.98). In 12/15 cases, the matched formalin-fixed and frozen samples predicted the same tumour molecular subtypes with confidence. These results demonstrate that the whole genome DASL assay is a valuable tool to profile degraded RNA from archival FFPE material. This assay will enable transcriptomic analysis of a large number of archival samples that are stored in pathology archives around the globe and consequently will have the potential to improve our understanding and characterization of many diseases.
组织样本采集是许多研究中的一个限制步骤。全世界有数千个福尔马林固定、石蜡包埋的存档块,但相比之下,肿瘤库中只有相对较少的新鲜冷冻样本。一旦样本在福尔马林固定后,RNA 就会降解,传统的基因表达谱分析方法就不适用了。在这项研究中,我们评估了 Illumina 的全基因组 DASL(cDNA 介导的退火、选择、延伸和连接)检测法在分析福尔马林固定、石蜡包埋(FFPE)存档乳腺肿瘤组织中的转录组的能力。我们对 76 个携带 BRCA1、BRCA2 或 ATM 突变的家族性乳腺癌病例或非 BRCA1/2 家族的病例进行了分析。我们发现,重复样本之间相关性良好(r(2) = 0.9-0.98)。在 12/15 例中,匹配的福尔马林固定和冷冻样本以信心预测了相同的肿瘤分子亚型。这些结果表明,全基因组 DASL 检测法是一种从存档的 FFPE 材料中分析降解 RNA 的有价值的工具。该检测法将能够对全球病理档案中储存的大量存档样本进行转录组分析,从而有可能提高我们对许多疾病的理解和特征描述。