Robust global micro-RNA profiling with formalin-fixed paraffin-embedded breast cancer tissues
Robust global micro-RNA profiling with formalin-fixed paraffin-embedded breast cancer tissues
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DOI：
10.1038/labinvest.2009.12
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发表时间：
2009-05-01
影响因子：
5
通讯作者：
Liu, Fei-Fei
中科院分区：
文献类型：
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作者：
Hui, Angela B. Y.;Shi, Wei;Liu, Fei-Fei
对人类恶性肿瘤进行的全球微小RNA（miR）分析日益增多，但到目前为止，大多数此类分析使用的是冷冻组织。然而，福尔马林固定是为了最佳保存细胞形态的标准常规组织学操作方法。为了确定对福尔马林固定组织进行miR分析是否可行，对40例存档的福尔马林固定石蜡包埋（FFPE）乳腺肿块切除术标本进行了miR表达的实时定量聚合酶链反应（qRT - PCR）分析。采用Taqman低密度芯片（TLDAs）评估365种miRs在34例浸润性导管癌以及6例来自乳房缩小整形术的正常对照中的表达水平。其技术重复性很高，样本内相关性高于0.9，在差异表达比较中的准确率为92.8%，这表明此类全局分析研究在技术和生物学上是可靠的。使用常规单孔qRT - PCR分析对TLDA数据进行了验证，结果显示这两种方法之间具有很强且具有统计学意义的一致性。来自同一患者的成对冷冻和FFPE乳腺癌样本显示出至少0.94的相似稳健相关性水平。与正常乳腺样本相比，一组miRs在乳腺癌中持续失调，包括先前报道的与乳腺癌相关的miRs，如上调的miR - 21、miR - 155、miR - 191和miR - 196a，以及下调的miR - 125b和miR - 221。还发现了其他可能具有生物学相关性的新型miR序列。这些结果表明使用FFPE样本进行全球miR分析的有效性和实用性，从而为评估与临床数据库相关联的此类材料的存档库提供了巨大机会，以便快速深入了解miRs在人类恶性肿瘤中的临床相关作用。
Global micro-RNA (miR) profiling of human malignancies is increasingly performed, but to date, the majority of such analyses have used frozen tissues. However, formalin fixation is the standard and routine histological practice for optimal preservation of cellular morphology. To determine whether miR analysis of formalin-fixed tissues is feasible, quantitative real-time PCR (qRT-PCR) profiling of miR expression in 40 archival formalin-fixed paraffin-embedded (FFPE) breast lumpectomy specimens were performed. Taqman Low Density Arrays (TLDAs) were used to assess the expression level of 365 miRs in 34 invasive ductal carcinomas and in 6 normal comparators derived from reduction mammoplasties. Its technical reproducibility was high, with intra-sample correlations above 0.9 and with 92.8% accuracy in differential expression comparisons, indicating such global profiling studies to be technically and biologically robust. The TLDA data were confirmed using conventional single-well qRT-PCR analysis, showing a strong and statistically significant concordance between these two methods. Paired frozen and FFPE breast cancer samples from the same patients showed a similar level of robust correlation of at least 0.94. Compared with normal breast samples, a panel of miRs was consistently dysregulated in breast cancer, including earlier-reported breast cancer-related miRs, such as upregulated miR-21, miR-155, miR-191, and miR-196a, and downregulated miR-125b and miR-221. Additional novel miR sequences of potential biological relevance were also uncovered. These results show the validity and utility of conducting global miR profiling using FFPE samples, thereby offering enormous opportunities to evaluate archival banks of such materials, linked to clinical databases, to rapidly acquire greater insight into the clinically relevant role for miRs in human malignancies.