SSH方法对食管癌相关差异基因cDNA文库的建立及初步分析

CONSTRUCTING THE DIFFERENTIAL EXPRESSING GENES cDNA LIBRARY OF ESOPHAGEAL CANCER BY SUPPRESSION SUBTRACTIVE HYBRIDIZATION AND ANALYZING THE GENES CORRELATIVE WITH TUMORIGENESIS

目的建立河北省磁县具有家族史的食管癌组织与食管正常黏膜组织差异表达的cDNA消减文库,并初步筛选过度表达的食管癌差异基因。方法应用抑制性消减杂交技术(suppression subtractive hybridization,SSH)方法对10对食管癌组织及正常食管黏膜组织进行差异基因的消减、克隆、测序、鉴定。结果建立了食管癌差异表达片段cDNA消减文库;并进一步对初步筛选出的20个食管癌差异表达的基因进行测序和同源性分析,得出与肿瘤密切相关的基因10个,分别为锌指蛋白14、锌指蛋白66、肿瘤相关钙信号转运蛋白1,FMNL2基因、真核细胞转录延长因子1A1、肌小管相关蛋白6、Toll样受体10、次级淋巴趋化因子、细胞色素P450酶4F8、桥粒糖蛋白3。结论应用SSH技术成功建立食管癌差异表达基因cDNA消减文库,并获得10个在食管癌组织中过度表达的基因。

Objective To construct the subtractive cDNA library of esophageal cancer with family history and normal esophageal mucosa tissues in Ci county of Hebei Province, and screen the differential expressing genes of esophageal cancer. Methods Ten pairs of esophageal cancer and normal esophageal mucosa tissues were investaged by suppression subtractive hybridization （SSH） ,the differential expressing genes were subtracted,cloned,blast analysised and identified. Results The subtractive cDNA library of esophageal cancer were constructed,20 genes fragments of esophageal cancer were selected to be measured, 10 genes associating with tumorigenesis and progression were obtained： ZNF14 （zinc finger protein 14）, ZNF66 （zinc finger protein 66）, TACSTD1（tumor-associated calcium signal transducer 1）,FMNL2（homo sapiens formin-like 2）, EEF1Al（eukaryotic translation elongation factor 1 alpha 1）, MTMR6 （myotubularin related protein 6）,TLR10 （toll-like receptor 10）,SLC31Al（solute carrier family 31 ,member 1）,CYP4F8 （cytochrome P450,family 4, subfamily F, polypeptide 8）, DSC3 （desmocollin 3）. Conclusion The subtractive cDNA library of esophageal cancer were obtained by SSH,and 10 genes that expressed in esophageal cancer were found.