Integrated Genetic Analysis of Allelic Imbalance and FGFR3 Mutation by SNP-based Pyrosequencing in Urothelial Cancer

基于 SNP 的焦磷酸测序对尿路上皮癌等位基因失衡和 FGFR3 突变进行综合遗传分析

• 批准号: 24890206
• 负责人: LUO Yi
• 金额: $ 1.08万
• 依托单位: Nara Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Research Activity Start-up
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-24890206/
• 关键词: 尿路上皮がん; 診断; 尿中膀胱癌診断; 遺伝子変化

In this study, we refer to the rapid and quantitative analytical methods in urine samples, a SNP-based pyrosequencing (PSQ) targeting regions of LOHs on 9p, 9q, 17p, and also exon 7, 10 which are hotspots of FGFR3 mutations. Material and Methods: A total of 7 markers were designed to amplify targeting SNP regions on 9p, 9q and 17p. And 2 markers were designed to validate FGFR3 sequence including 2 point mutated hotspot in each marker in exon 7 and 10. Results: A total of 116 UC samples are analyzed. In the tissue analysis, 85.0% of UC showed either LOH or FGFR3 mutation. In the analysis of urine sediments, 75.0% of urine obtained from UC patients’ showed genetic alterations. Urine cytology could detect only 44.0% of UCs in the same cohort. No genetic alterations were detected in 20 healthy urine sediments. Conclusions: PSQ were the feasible assay for detect genetic alterations such as LOH and point mutation from small cancer cell populations.

在本研究中，我们参考了尿液样本的快速和定量分析方法，基于SNP的焦磷酸测序（PSQ）针对9p，9q，17p上的LOH区域，以及外显子7，10，这是FGFR 3突变的热点。材料和方法：设计7个标记，分别扩增9p、9q和17p上的SNP区域。设计了2个标记来验证FGFR 3序列，每个标记在外显子7和10中包括2个点突变热点。结果：共分析了116份UC样本。在组织学分析中，85.0%的UC存在洛或FGFR 3突变。在尿沉渣分析中，75.0%的UC患者尿中存在基因改变。尿细胞学检查只能检出44.0%的UC。在20例健康人尿沉渣中未检测到遗传变异。结论：PSQ是检测小细胞群体中洛缺失和点突变等基因改变的可行方法。