Feasibility of using tissue microarray cores of paraffin-embedded breast cancer tissue for measurement of gene expression: a proof-of-concept study
Feasibility of using tissue microarray cores of paraffin-embedded breast cancer tissue for measurement of gene expression: a proof-of-concept study
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DOI：
10.1136/jcp.2010.075754
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发表时间：
2010-06-01
影响因子：
3.4
通讯作者：
Dowsett, Mitch
中科院分区：
文献类型：
--
作者：
Drury, Suzanne;Salter, Janine;Dowsett, Mitch
Aims To determine whether 0.6 mm core of formalin fixed paraffin embedded（FFPE）tissue，as commonly used to construct immunohistochemical tissue microarrays，may be a valid alternative to tissue section as source material for quantitative real-time PCR-based transcriptional profiling of breast cancer.Methods Four matched 0.6 mm core of invasive breast tumor and two 10 mu m whole section were taken from eight FFPE blocks.提取RNA并逆转录，并对Oncotype DX乳腺癌测定的21个基因进行TaqMan测定。16个复发相关基因的表达进行归一化的一组5个参考基因，和复发评分（RS）calculated.Results RNA产量低于从0.6毫米的核心比从10 μ m的整个部分，但仍然是足够的进行测定。RS和单基因数据的核心是高度可比的，从整个部分（RS p=0.005）。更大的变异性之间的核心比section.Conclusions之间FFPE部分是最好的0.6毫米核心RNA分析，以最大限度地提高RNA产量，并允许标准的组织病理学评估。然而，0.6 mm的芯就足够了，适合用于大型队列研究。
Aims To determine whether 0.6 mm cores of formalin-fixed paraffin-embedded (FFPE) tissue, as commonly used to construct immunohistochemical tissue microarrays, may be a valid alternative to tissue sections as source material for quantitative real-time PCR-based transcriptional profiling of breast cancer.Methods Four matched 0.6 mm cores of invasive breast tumour and two 10 mu m whole sections were taken from eight FFPE blocks. RNA was extracted and reverse transcribed, and TaqMan (R) assays were performed on the 21 genes of the Oncotype DX (R) Breast Cancer assay. Expression of the 16 recurrence-related genes was normalised to the set of five reference genes, and the recurrence score (RS) was calculated.Results RNA yield was lower from 0.6 mm cores than from 10 mu m whole sections, but was still more than sufficient to perform the assay. RS and single gene data from cores were highly comparable with those from whole sections (RS p=0.005). Greater variability was seen between cores than between sections.Conclusions FFPE sections are preferable to 0.6 mm cores for RNA profiling in order to maximise RNA yield and to allow for standard histopathological assessment. However, 0.6 mm cores are sufficient and would be appropriate to use for large cohort studies.