摘要
目的:胃黏膜异型增生是一种公认的重要癌前病变,筛查癌前病变相关基因并研究这些基因在胃癌不同阶段的表达,探讨胃癌发生分子机制.方法:手工显微切割取胃异型增生和正常组织,应用cDNAPCR方法对少量组织全基因组扩增后进行双向抑制性消减杂交,消减后片段与载体连接、克隆、筛选、测序及同源性检索.应用斑点杂交检测基因在胃癌不同阶段的表达,并用半定量RT-PCR方法进一步验证检测结果.结果:正常和异型增生组织互为tester和driver成功构建了两个cDNA消减文库,测序的26个克隆中21个片段在胃癌不同阶段有表达异常,特别是其中4个基因(P125,cytochromecoxidasesubunitI,meprinA,acidiccalponin)在异型增生、早癌、进展期胃癌中皆有表达改变,可能是重要的胃癌癌前病变相关基因.结论:发现4个新的与胃癌发生相关基因,其具体机制有待进一步研究.
AIM:To explore molecular mechanism of gastric carcinogenesis, we screened associated genes of gastric dysplasia and further investigated their expression in gas- tric carcinomas with different stages. METHODS:Relatively pure dysplasia and normal tissue were procured by manual microdissection, amplified by cDNA- PCR, and then used to carry out forward (dysplasia as tester, normal tissue as driver) and reverse (normal tissue as tester, dysplasia as driver) SSH. Subtracted cDNA fragments were cloned into vector, screened, sequenced, and made homolo- gous analysis. The expression of differentially expressed fragments was detected and verified by Dot hybridization and reverse transcription-PCR. RESULTS:Two subtracted cDNA libraries were constructed. Twenty-one of 26 sequenced clones were verified to be ex- pressed differentially. It was noted that differentiall expres- sions of 4 genes (P125,cytochrome c oxidase subunit I, meprin A,acidic calponin)were detected simultaneously in dysplasia, early cancer and advanced cancer.CONCLUSION:Four new associated genes have been iden-tified in dysplasia. Further studies are necessary to deter- mine their roles in gastric carcinogenesis.
出处
《世界华人消化杂志》
CAS
2003年第1期6-9,共4页
World Chinese Journal of Digestology
基金
国家重点基础研究发展规划项目(973项目)
G1998051203~~
