食管鳞癌中神经纤毛蛋白2基因断裂和重排的分析与鉴定  

Identification of genetic disruptions and rearrangement of neuropilin 2 in esophageal squamous cell carcinoma

作　　者：程潇钰 郝佳洁[1] 宫婷[1] 商利[1] 蔡岩[1] 张钰[1] 徐昕[1] 王明荣[1] Cheng Xiaoyu Hao Jiafie Gong Ting Shang Li Cai Yan Zhang Yu Xu Xin Wang Mingrong(State Key Laboratory of Molecular Oncology, National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China)

机构地区：[1]国家癌症中心/中国医学科学院北京协和医学院肿瘤医院分子肿瘤学国家重点实验室,北京100021

出　　处：《国际遗传学杂志》2016年第5期233-239,共7页International Journal of Genetics

基　　金：国家自然科学基金（81330052、81321091）

摘　　要：目的分析食管鳞癌中的基因断裂及重排，旨在为食管鳞癌的诊断及治疗提供分子标志和新的治疗靶点。方法通过对食管鳞癌手术标本进行单核苷酸多态性（singlenucleotide polymorphism，SNP）微阵列检测，首先发现基因内部断裂；采用cDNA末端快速扩增法（rapid amplification of cDNAends，RACE）技术鉴定与断裂基因残留部分融合的新的染色体片段，采用蛋白免疫印迹（Westernblotting）和免疫组织化学（immunohistochemistry，IHC）方法检测基因断裂后的表达情况；利用荧光原位杂交（fluorescence in situ hybridization，FISH）技术检测候选基因在较大临床样本中的断裂频率。结果SNP微阵列分析6例食管鳞癌手术组织，在2例标本中发现2号染色体长臂33．3区段的MRP2基因内发生了断裂。3’-RACE分析结果表明，在其中1例的互补DNA水平存在神经纤毛蛋白2-膜联蛋白A13基因（NRP2-ANXA，3）融合转录本。通过FISH进一步分析16例食管鳞癌组织，发现6例存在NRP2断裂，其中2例疑似存在NRP2-ANXAJ3重排。Western印迹和IHC分析结果显示，发生NRP2-ANXA13融合病例标本的正常NRP2蛋白表达与癌旁组织相比显著下调。结论在食管鳞癌组织中存在NRP2基因的高频断裂和NRP2-ANXA13的基因融合，该结果为探明食管鳞癌中基因断裂和重排的作用及机制提供了新的重要线索。Objective Research on genetic disruptions and rearrangements in esophageal squa- mous cell carcinoma （ESCC） may provide new targets in prognostic and clinical treatments. The present study is to identify genetic disruptions and rearrangements in ESCC samples. Methods Six operative sam- ples were analyzed by SNP microarrays to identify genetic disruptions in ESCC. RACE experiment was car- ried out to find new fusion transcripts. Fluorescence in situ hybridization （FISH） was applied for screen- ing of the frequency of genetic disruptions. Western blotting and immunohistochemistry （ 1HC ） were taken to detect the expression of the splicing genes in ESCC samples. Results Genetic disruptions of NRP2 on chromosome 2q33.3 were found by SNP micro-array in two out of six ESCC samples. NRP2-ANXA13fusion transcripts were found using 3＇-RACE experiment in one sample harbouring NRP2 disruption. FISH results indicated that high frequency of NRP2 disruptions existed in 37. 5 % （6/16） ESCC samples while NRP2-ANXA13 likely existed in two out of them. These Western blotting and IHC results indicate that NRP2 was down-regulated in the sample harboring NRP2-ANXA13 which was detected by SNP microar- ray. Conclusion NRP2 disruptions and NRP2-ANXA13 were identified in ESCC for the first time in our experiment, which provides an important clue in exploring the significance of genetic disruptions and rear- rangements in ESCC.

关 键 词：NRP2 基因断裂 基因融合 

分 类 号：R735.1[医药卫生—肿瘤]