人食管癌相关基因cDNA片段的克隆、筛选与初步鉴定  

Cloning and identification of human esophageal cancer associated gene cDNA fragments

作　　者：陈东[1] 张云汉[1] 殷智榕[1] 高冬玲[1] 姜国忠[1] 贺建业[2] 王尧河[1] 寿思明[1] 

机构地区：[1]河南医科大学第一附属医院病理科,河南省肿瘤病理重点实验室,郑州450052 [2]郑州铁路局中心医院普外科,郑州450052

出　　处：《河南医科大学学报》2001年第5期526-530,共5页Journal of Henan Medical University

基　　金：河南省科学技术发展计划资助项目 ( 97 6 2 0 )

摘　　要：目的 :筛选和鉴定人原发性食管癌组织的相关基因 ,揭示食管癌的癌变机理。方法 :用高效、灵敏的荧光mRNA差异显示技术 ,以食管癌及相应的正常食管粘膜组织为对照 ,通过对其基因表达的比较 ,找出差异条带 ,利用ReverseNorthernDotBlot、DNA序列分析和NorthernBlot,并结合mRNA原位杂交技术对被筛片段进行鉴定。结果 :①在实验中分离、鉴定了 74条差异片段 ,其中包括正常组织表达而癌组织中不表达的差异片段 5 4个 ,癌组织中表达而正常组织不表达的差异片段 2 0个。②其中 1个片段C5 7(337bp)在GenBank数据库中没有发现同源的已知基因或片段 ,推测其可能为食管癌相关基因。③经NorthernBlot检测C5 7在癌组织中有表达信号。④原位杂交技术检测C5 7在癌组织中具有较高的阳性表达率 (74% )。结论 :食管癌组织中C5 7可能是新的食管癌相关的候选癌基因 。Aim: To screen and identify new genes related to human primary esophageal cancer in order to reveal the mechanism of carcinogenesis.Methods: We isolated the differential fragments of oncogenes and tumor suppression genes by using sensitive non radioactive isotope fluorescence labeled mRNA differential display technique from esophageal cancer and matched adjacent normal mocusa tissues from surgical resection in the same patients. Then reverse northern dot blot, northern blot, homology analysis and in situ hybridization technique were applied to identify these fragments. Results: ① Seventy four cDNA fragments were separated ranging from 300 to 900 bp,and 20 differential fragments were highly expressed in cancer tissues that was named 'C', and the other 54 fragments were highly expressed in normal mucosa tissues named 'N'. ②Among the 74 cDNA fragmeuts,C57 fragment was not homologous to the known sequences in the Advanced Blast database of GenBank,and was assigned as esophageal cancer associated gene. ③Northern blot hybridization showed signal of C57 expression in cancer tissues. ④ISH technique showed that the expression of C57 between normal epithelial and esophageal cancer tissues was significantly different. Conclusions: Fragment C57 may contribute to the causation and progression of esophageal cancer, and may be regarded as one of new candidates of oncogenes.

关 键 词：食管癌 MRNA差异显示技术 杂交 CDNA 

分 类 号：R735.1[医药卫生—肿瘤]