大鼠纤维肉瘤模型中等位基因失衡的微卫星标记分析

Å. Sjöling, A. Walentinsson, G. Levan, K. Klinga-Levan
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引用次数: 1

摘要

采用皮下注射DMBA(7,12-二甲基苯[a]-蒽)诱导大鼠近交系BN/Han与LE/Mol杂交后代发生纤维肉瘤。34只接受治疗的动物中有21只(62%)在注射部位发生了纤维肉瘤。分析肿瘤DNA的等位基因失衡,这将表明遗传物质的扩增或缺失。为了覆盖整个大鼠连锁图谱的长度,包括所有20个常染色体和X染色体,我们选择了215个多态性微卫星标记。标记物每组6-12个标记,经荧光PCR扩增后可在聚丙烯酰胺凝胶上同时分析。利用纤维肉瘤材料上的面板揭示了许多等位基因不平衡的病例。其中大多数涉及位于大鼠1号和2号染色体上的标记,表明位于这些染色体上的基因在肉瘤发生中起重要作用。这些微卫星标记面板的发展将极大地促进未来通过全基因组扫描对大鼠肿瘤进行分析。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Microsatellite marker analysis of allelic imbalance in a rat fibrosarcoma model

Fibrosarcomas were induced by subcutaneous injections of DMBA (7,12-dimethylbenz[a]-anthracene) in progeny from a cross between the inbred rat strains BN/Han and LE/Mol. Twenty-one of 34 treated animals (62%) developed fibrosarcomas at the injection site. The tumor DNA was analyzed for allelic imbalance that would be indicative of amplification or deletion of genetic material. In order to cover the entire length of the rat linkage map, including all 20 autosomes and the X chromosome, 215 polymorphic microsatellite markers were selected. The markers were grouped into panels of 6–12 markers, which could be analyzed simultaneously on polyacrylamide gels after fluorescent PCR amplification. Numerous cases of allelic imbalance were revealed using the panels on the fibrosarcoma material. Most of them involved markers situated on rat chromosome 1 and 2, suggesting that genes located on these chromosomes are important in sarcomagenesis. The development of microsatellite marker panels such as those described will greatly facilitate future analysis of rat tumors by genome-wide scanning.

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