Multiplex genotype analysis of invasive carcinoma and accompanying proliferative lesions microdissected from breast tissue.
Multiplex genotype analysis of invasive carcinoma and accompanying proliferative lesions microdissected from breast tissue.
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从乳腺组织显微解剖的浸润性癌和伴随的增殖性病变的多重基因型分析。
DOI：
10.1016/s1525-1578(10)60612-5
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Li,H
中科院分区：
文献类型：
--
作者：
Cui,X;Feiner,H;Lin,Z;Li,H
为了全面了解乳腺癌的遗传基础，需要在大量的遗传标记位点上分析所谓的前驱病变，并相互比较，并与侵袭性成分进行比较。然而，大多数这些病变的显微尺寸和通过显微切割可以获得的非常少量的材料限制了可以包括在分析中的位点的数量。为了解决这个问题，已经开发了多重基因分型方法。用这种方法，在28个标记位点的多态性序列同时从5 μm石蜡包埋的乳腺组织切片的显微切割的组成部分扩增。通过变性梯度凝胶电泳解析扩增的等位基因产物后确定病变的基因型。由于从单个5 μm切片中的每个病变中分离的材料足以用于几个28个位点的测定，并且可以制备具有相同病变组的几个连续组织切片，因此可以确定每个病变在数百个遗传标记位点上的基因型，这些遗传标记位点可以很好地覆盖人类基因组。分析足够数量的病例可能会产生可用于全面了解乳腺癌发展遗传基础的信息。
To understand the genetic basis of breast cancer in a comprehensive way, purported precursor lesions need to be analyzed at a large number of genetic marker loci and compared with each other and with the invasive components. However, the microscopic size of most of these lesions and the very small amount of material that can be obtained through microdissection limit the number of loci that can be included in the analysis. To address this issue, a multiplex genotyping approach has been developed. With this approach, polymorphic sequences at 28 marker loci were amplified simultaneously from the microdissected components in 5-μm paraffin-embedded breast tissue sections. The genotypes of the lesions were determined after resolving the amplified allelic products by denaturing gradient gel electrophoresis. Because the material isolated from each lesion in a single 5-μm section was sufficient for several 28-locus assays and several successive tissue sections with the same set of lesions may be prepared, it is possible to determine the genotype of each lesion at hundreds of genetic marker loci that may well cover the human genome. Analyzing a sufficient number of cases may yield information that could be used to understand the genetic basis of breast cancer development in a comprehensive way.