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

在本研究中，我们参考了尿液样本的快速定量分析方法，基于snp的焦磷酸测序（PSQ）靶向9p， 9q， 17p以及外显子7,10上的LOHs区域，这些区域是FGFR3突变的热点。材料与方法：共设计7个标记，针对9p、9q和17p上的SNP区域进行扩增。设计了2个标记来验证FGFR3序列，每个标记在外显子7和10上都有2个点突变热点。结果：共分析了116例UC样本。在组织分析中，85.0%的UC显示LOH或FGFR3突变。在尿液沉积物分析中，UC患者尿液中有75.0%显示遗传改变。尿细胞学检查在同一队列中仅能检出44.0%的UCs。20例健康尿液沉积物未检测到基因改变。结论：PSQ是检测小肿瘤细胞群LOH和点突变等基因改变的可行方法。

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.