Combined laser capture microdissection and serial analysis of gene expression from human tissue samples
Combined laser capture microdissection and serial analysis of gene expression from human tissue samples
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DOI：
10.1038/modpathol.3800327
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发表时间：
2005-04-01
期刊：
影响因子：
7.5
通讯作者：
McDonnell, TJ
中科院分区：
文献类型：
--
作者：
Cho-Vega, JH;Troncoso, P;McDonnell, TJ
来自异质性人类组织的细胞特异性基因表达谱受到细胞纯化限制的混淆。在这里，我们描述了一种技术，以产生从新鲜冷冻前列腺切除术标本和小的初始数量的RNA相结合的激光捕获显微切割和基因表达的微序列分析（LCM-microSAGE）的前列腺癌细胞的纯群体的基因表达谱。从大约100000个激光脉冲中获得了两个microSAGE文库，估计含有少于3 × 10（5）个细胞和20-30 ng mRNA。两个文库测序的深度为10 111和10 463个独特的标签从正常和癌细胞，分别代表6453和6923基因。大多数转录物以相似的水平表达，但与正常细胞相比，癌细胞的385个标签的表达增加，389个标签的表达减少。共发现20个差异表达基因（P
Cell-specific gene expression profiling from heterogeneous human tissues is confounded by cell purification limitations. Here, we describe a technique to generate gene expression profiles of pure populations of prostate cancer cells obtained from fresh-frozen prostatectomy specimens and small initial quantities of RNA by combining laser capture microdissection and microserial analysis of gene expression (LCM-microSAGE). Two microSAGE libraries were obtained from approximately 100 000 laser pulses, estimated to contain fewer than 3 x 10(5) cells and 20-30 ng mRNA. Two libraries were sequenced to a depth of 10 111 and 10 463 unique tags from normal and cancer cells, representing 6453 and 6923 genes, respectively. Most transcripts were expressed at similar levels, but cancer cells compared with normal cells had increased expression of 385 tags and decreased expression of 389 tags. A total of 20 genes were differentially expressed (P