银染mRNA差异显示法的建立及其应用  

A Modified Silver Stained Differential DisplayMethod and Its Application

作　　者：陈瑞川[1] 蔡克瑕[1] 马胜平[1] 陈福[1] 付永贵[1] 曾寰[2] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005 [2]厦门市第二医院肿瘤实验室,福建厦门361003

出　　处：《厦门大学学报（自然科学版）》2001年第5期1138-1144,共7页Journal of Xiamen University：Natural Science

基　　金：福建省自然科学基金资助项目 (F0 0 0 2 4 )

摘　　要：以银染 m RNA差异显示方法对原发及转移灶胃癌标本总 RNA为研究对象进行银染差示分析和回收差异条带 ,从中获得系列差异表达片段 .随机选取 5个从原发性胃癌样品回收的表达条带 ,以取自体外培养胃癌细胞株的 RNA进行点杂交验证 ,均被证实为真实带 .对 2个差异表达片段进行分析、测序和 Gen Bank同源比较结果表明 :MGD1片段与肿瘤转移相关基因 MTA1完全同源 ,属胃癌转移相关基因 ;PGD2与胃癌转移抑制相关 ,并与细胞周期抑制蛋白 p2 7/ Kip1具 99%的同源性 .结果表明 ,银染差异显示法具有快速、直观、假阳性率低等优点 。Described here was a silver stained mRNA differential display PCR(DD PCR)meth od which was modified from traditional DD PCR technique and used for analysis o f differential expressed genes between primary and metastatic gastric can cer tissues. To confirm the reliability of this method, five PCR fragments were randomly selected from primary gastr ic cancer sample and were verified by RNA dot blot with total RNA extracted from cultured gastric ca ncer cell lines. Two fragmentsMGD1 and PGD2 were subjected to verificatio n by RNA dot blot with total RNA respectively extracted from primary and metastat ic gastric cancer tissues and final sequencing. The confirmation results of five fragments showed that the randomly selected five fragments were all real produc ts , indicating that the silver stained DD PCR method possesses a good reliability . The analysis results of two fragments showed that MGD1, highly expressed in me tastatic gastric cancer, is homology to MTA1 (metastasis associated gene 1), an d PGD2, highly expressed in primary gastric cancer, is 99% homology to cell cycl e inhibitor p27/Kip1 gene, which may provide the new clues for research of molec ular mechanism of gastric cancer metastasis.

关 键 词：银染差示PCR 胃癌 转移相关基因 银染mRNA差异显示法 肿瘤转移 差异表达片段 

分 类 号：R735.2[医药卫生—肿瘤] R730.49[医药卫生—临床医学]