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激光捕获显微切割技术在神经母细胞瘤及神经母细胞瘤前体细胞遗传分析中的应用。

Application of laser capture microdissection in genetic analysis of neuroblastoma and neuroblastoma precursor cells.

作者信息

De Preter Katleen, Vandesompele Jo, Heimann Pierre, Kockx Mark M, Van Gele Mireille, Hoebeeck Jasmien, De Smet Els, Demarche Martine, Laureys Geneviève, Van Roy Nadine, De Paepe Anne, Speleman Frank

机构信息

Department of Medical Genetics, Ghent University Hospital, 1K5, De Pintelaan 185, B-9000 Ghent, Belgium.

出版信息

Cancer Lett. 2003 Jul 18;197(1-2):53-61. doi: 10.1016/s0304-3835(03)00084-3.

Abstract

Recently developed quantitative and high-throughput technologies that allow automated and rapid screening of the whole genome, transcriptome and proteome have revolutionized the field of cancer genetics. At the same time, new challenges are met, e.g. the need for improved data analysis and standardization of tumor sample handling. Even if these issues are resolved, an 'old' problem in genetic tumor analysis remains, i.e. contamination of tumor samples by stromal and surrounding normal cells. To overcome this obstacle, laser capture microdissection (LCM) has been developed in order to procure the cells of interest from stained tissue sections with retention of morphology. In this review we describe the possible down-stream applications of LCM in the genetic analysis of neuroblastoma (NB). Special focus is given to MYCN copy number determination using real-time quantitative polymerase chain reaction (Q-PCR), analysis of 1p-, 3p- and 11q-deletions using loss of heterozygosity analysis and Q-PCR expression analysis of microdissected normal neuroblast cells and NB cells.

摘要

最近开发的定量和高通量技术能够对全基因组、转录组和蛋白质组进行自动化快速筛选,彻底改变了癌症遗传学领域。与此同时,也面临着新的挑战,例如需要改进数据分析以及规范肿瘤样本处理。即便这些问题得到解决,遗传肿瘤分析中的一个“老”问题依然存在,即肿瘤样本被基质细胞和周围正常细胞污染。为克服这一障碍,人们开发了激光捕获显微切割技术(LCM),以便从染色的组织切片中获取感兴趣的细胞并保留其形态。在本综述中,我们描述了LCM在神经母细胞瘤(NB)遗传分析中可能的下游应用。特别关注使用实时定量聚合酶链反应(Q-PCR)测定MYCN拷贝数、使用杂合性缺失分析以及对显微切割的正常神经母细胞和NB细胞进行Q-PCR表达分析来检测1p、3p和11q缺失。

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