摘要
目的:应用甲基化敏感性代表性差异分析法(methylation-sensitive representational difference analysis,MS-RDA)筛选胃癌和正常胃组织间甲基化差异DNA片段．方法:通过MS-RDA筛选胃癌和正常胃组织间DNA甲基化差异片段,经克隆、测序后进行生物信息学分析．结果:获得3个甲基化差异片段,分别为 CRS1308,CRS1309,CRS1310k列,其中 CRS1309和CRS1310已被GenBank收录,登陆亏分别为AY887106和AY887107,CRS1309 序列与LOC440683基因第11外显子、 LOC440887基因的3’端,DRD5基因启动子和外显子区域均有很高的相似性(分别为98％,99％, 94％),CRS1310序列与1999年Minoru Toyota在人类结肠癌中分离出的核糖体RNA上的甲基化差异性CpG岛有很高的相似性(98％)．结论:胃癌和正常胃组织间DNA甲基化存在差异,MS-RDA可有效分析这两种不同组织间甲基化的差异,筛选出有意义的甲基化差异片段.
AIM： To isolate the differentially methylated DNA sequences between gastric cancer and normal gastric mucosa. METHODS： The differentially methylated DNA sequences between gasrtic cancer and normal gastric mucosa were isolated by methylationsensitive representational difference analysis （MS-RDA）. Similarities between the separated fragments and the human genomic DNA were analyzed with Building Loads Analysis and Sytem Thermodynamics （BLAST） program. RESULTS： Three differentially methylated DNA sequences, CRS1308, CRS1309, and CRS1310, were obtained. CRS1309 and CRS1310 had been accepted by GenBank. CRS1309 was highly similar to the eleventh exon of LOC440683 （98%）, 3′ end of LOC440887 （99%）, and promoter and exon region of DRD5 （94%）. CRS1310 was consistent （98%） with a CpG island, in ribosomal RNA, isolated from colorectal cancer by Minoru Toyota in 1999. CONCLUSION： The methylation degree is different between gastric cancer and normal gastric mucosa. The differentially methylated DNA sequences can be isolated effectively by MS-RDA.
出处
《世界华人消化杂志》
CAS
北大核心
2006年第4期365-369,共5页
World Chinese Journal of Digestology
基金
湖南省科研计划重大专项
No.04SK1004
湖南省教育厅科学研究资助项目.No.99C195
