胃癌高表达cDNA片段W41的克隆及组织表达谱分析

Cloning and tissue expression analysis of up-regulated cDNA fragment in human gastric cancer

目的 从人胃癌组织中克隆新的相关易感基因。方法 利用 c DNA末端快速扩增 PCR技术得到了扩增片段 ,将其克隆、核苷酸序列分析 ,并将序列在 Gen Bank进行同源性比较。利用 Northern杂交、多组织 Northern及基因表达系列分析了所得基因片段的组织表达谱。结果 获得 1条 5 33bp带有 poly(A)尾的 c DNA片段 ,可见加尾信号 AATAAA。与 Gen Bank基因数据库同源比较 ,该序列未见与任何已知基因同源 ,登录 Gen Bank(登录号为 AF32 5 2 0 2 )。该序列在胃癌组织的表达强度高于对应正常组织 ,多组织Northern及基因表达系列分析表明该序列在多种肿瘤组织中高表达 ,在正常组织中表达减弱。结论 得到了 1条与胃癌可能相关的新的 c

Objective: To identify novel human gastric cancer-associated susceptibility gene for early diagnosis and treatment of gastric cancer. Methods: A primer was designed for 3′-rapid amplification of cDNA end (RACE) and amplified fragments were cloned, then they were analyzed by sequencing. Compared with ESTs in Genbank, the EST fragment represented a novel gene. Combination of Northern blot and virtual Northern and multiple tissues Northern blot, expression of the cDNA in multiple normal and carcinoma tissues were analyzed. Results: One of the important cDNA bands with poly(A) tail was cloned. This band was named W41. Sequence analysis showed that W41 consists of 533 bp. Basic local alignment search tool analysis revealed that W41 has low identity with any genes from GenBank. This sequence data was submitted to GenBank with accession No. AF 325202. Northern blot revealed that W41 presented higher expression in gastric cancer tissue than in normal tissue. Multiple tissue Northern blot revealed that W41 presented higher expression in mutiple cancers than in normal tissues. Virtual Northern revealed that the cDNA presented higher expression in tumor series analysis of gene expression libraries than in normal. Conclusion: A novel human gastric cancer-associated cDNA fragment was identified.