应用全外显子测序和显微切割技术识别1例肺癌患者高频突变基因的异质性探索

Using Whole-Exome Sequencing and Laser Capture Microdissection to Recognize High Mutation Genes and Investigate the Heterogeneity in one Patient with Lung Cancer

目的:通过对一例肺鳞癌患者全外显子测序来识别这例肺癌的可能致病基因,并通过显微切割初步探索这例肺癌肿瘤细胞的起源与演化。方法:利用全外显子测序技术对肺癌肿瘤组织和相应癌旁组织测序;用COSMIC肿瘤数据库比较分析统计出肺癌可能致病基因;用激光显微切割技术提取五个不同部位肿瘤细胞;巢式PCR扩增,一代测序验证基因分型。结果:发现了这例肺癌病人的7个高频突变基因:LPHN2、TP53、MYH2、TGM2、C10orf137、MS4A3和EP300;这些基因在10×镜下和20×镜下经显微切割的肺癌组织的5个不同部位上的基因分型不同。结论:我们通过全外显子测序发现了这例肺癌的7个可能致病基因,并初步探索了这例肺癌肿瘤细胞是多克隆起源的。

Objective： To discover suspicious driver genes of one lung squamous carcinoma patient by Whole-Exome Sequencing and to preliminarily investigate the origin and progression of this cancer through laser capture microdissection. Methods： We conducted the Whole-Exome Sequencing of a lung squamous carcinoma and adjacent normal tissue; compared with COSMIC tumor database, we inferred several suspicious driver genes of lung cancer; tumor cells from five different directions were microdissectedbylaser capture microdissection; the genome typing was determined by nest- polymerase chain reaction （nest-PCR） and Sanger sequencing. Results： We inferred seven high mutation genes of lung cancer： LPHN2, TP53, MYH2, TGM2, C 10orf137, MS4A3 and EP300; the genome typing of these seven genes were distincted in five dissections both in 10× and 20× microscope. Conclusions： We inferred seven suspicious driver genes of this lung squamous carcinoma by Whole-Exome Sequencing, and preliminarily discovered that this lung cancer cell may be derived from polyclonal cells.