摘要
目的:应用荧光mRNA差异显示技术(DD-PCR),筛选胃癌及食管癌相关基因的异常表达。方法:收集临床胃癌组织样本,通过荧光差异显示技术(DD-PCR)获得胃癌样品中差异的片段,对这些片段进行克隆和测序。通过在GenBank中同源性检索,查找与差异片段相对应的同源基因。利用半定量PCR及定量PCR方法检验该基因在胃癌及食管癌组织中与其对应正常组织之间表达的差异。结果:通过DD-PCR获得的一个差异表达片段的序列对应于ATP/GTP结合蛋白1基因(A/GT-PBP1)。半定量PCR及荧光定量PCR技术检测结果表明,A/GTPBP1基因在胃癌及食管癌组织中的表达量高于其对应的正常组织(胃癌,P<0.01;食管癌,P<0.01)。该基因包含25个外显子,读码框长3561bp,编码1186个氨基酸,蛋白质相似性分析表明该蛋白为G蛋白家族成员。结论:A/GTPBP1在胃癌组织中异常表达,可能在胃癌发生过程中起调节作用。
Objective： The mRNA differential display （DD-PCR） was employed to search genes expressed differently in gastric cancer tissues. Methods：Gastric carcinoma tissues collected from clinical sample in hospital, mRNA differential display （DD-PCR） was employed to search differently expressed fragments, some of which were cloned and sequenced. By homology analysis in GenBank, corresponding homologous genes of those fragments were found. The corresponding genes of those differently expressed fragments were validated by semi-real time and real-time quantitative PCR. Results：By DD PCR, one EST was identified to be ATP/GTP binding protein 1 gene （A/GTPBP1）. The result of semi-real time and real-time quantitative PCR showed that the expression level of ATP/GTP binding protein 1 gene both in gastric cancer （P〈0.01） and esophageal carcinoma （P〈0.01） was significantly higher than that in normal tissues. The ORF of ATP/GTP binding protein 1 gene was a561 base pair long and encodes 1186 amino acids. The molecular weight was about 84.4KD. Conclusion： The over-expression of ATP/GTP binding protein 1 gene in gastric and esophageal cancer tissues may play an important regulation role in the improvement of gastric and esophageal carcinogenesis.
出处
《中国临床医学》
北大核心
2008年第2期186-188,共3页
Chinese Journal of Clinical Medicine
基金
江苏省社会发展基金资助项目(BS2006041)
关键词
荧光差异显示
胃癌
食管癌
ATP/GTP结合蛋白1
定量PCR
Fluorescent differential display（ FDD）
Gastric cancer
Esophageal cancer
ATP/GTP binding protein 1~ Real-time quantitative PCR
