应用mRNA差异显示技术筛选子宫内膜癌相关基因片段

Application of mRNA differential display technique in screening related gene of endometrial carcinoma

目的 应用mRNA差异显示技术 ,筛选子宫内膜癌的相关基因片段。方法 提取同一患者的正常子宫内膜和子宫内膜癌组织总RNA ,行差异显示 聚合酶链反应 (DD PCR)技术 ,扩增产物在测序胶上电泳 ,经银染分离筛选差异条带并测序 ,用BLAST基因数据库进行分析。结果 有 2条基因片段与已知人类基因片段同源性低或无同源性 ,自定义为T1 7,T1 7基因片段登陆号为AF315 5 81;另 2条基因片段与已知基因片段同源性高达 97 3%和 85 0 %。T1 7基因片段在子宫内膜癌组织中呈高表达 ,在增生的子宫内膜组织中呈低表达 ,在正常子宫内膜组织中不表达。结论 T1 7基因片段可能为子宫内膜癌新的相关基因片段。

Objective To screen related gene of endometrial cancer by mRNA differential display (DD) polymerase chain reaction (PCR). Methods Individual mRNA from normal and neoplastic endometrium obtained from the same patient were comparatively studied by DD methodology.The mRNAs were isolated from the tissues, reverse transcribed to cDNA, and then amplified by PCR. The PCR products were displayed on a polyacrylamid gel by silver staining, and the differentially expressed bands were retrieved and reamplified. Subsequently we selected four bands for sequencing and making further investigation. Results cDNA homology was analysed by searching through GenBank sequence databases and indicated that tumor fragment (T1 7) has no significant sequence homology with any sequence of known function. Acession number is AF315581. Further study showed that T1 7 expressed more abundantly in endometrial cancer than hyperplasia endometrium, but not expressed in normal endometrium. Conclusions T1.7 is a new expressed sequence tag. It may be related gene fragment of endometrial cancer.