激光捕获显微切割与荧光差异显示技术结合应用于子宫内膜癌相关基因的研究

Application of laser capture microdissection combined with fluoro differential display in the research of the relevant genes of endometrial carcinoma

目的:排除混杂细胞的干扰,获取并研究人子宫内膜癌发病特异相关基因,探索子宫内膜癌发生的分子机制。方法:运用激光捕获显微切割技术(LCM)获取无间质混杂的人正常子宫内膜腺管上皮细胞与子宫内膜癌细胞,提取微量RNA并进行纯化及浓缩,采用荧光差异显示技术(FDD-PCR),筛选与子宫内膜癌发病特异相关的基因片段,克隆测序并进行分析。结果:经杂交鉴定获得6条差异基因片段,1条在正常子宫内膜中高表达,5条在子宫内膜癌中高表达。结论:通过LCM技术与FDDRT-PCR技术的结合,获得了与子宫内膜癌发病特异相关的基因片段。CDK-7等基因可能与子宫内膜癌的发生发展有密切的关系。

OBJECTIVE： To exclude the disturb of mixed cell types, acquire human endometrial carcinoma special relevant genes, and explore the molecular basis of cancer genesis. METHODS：Pure endometrial glandular epithelial cells and endometrial carcinoma cells were obtained by LCM, RNA from these cells was isolated, and genes that were expressed differentially were identified with the fluoro differential display reverse transcription polymerase chain reaction （FDD-PCR）, and the selected fragments were cloned, sequenced and analysed. RESULTS： Confirmed by Reverse Northern Blot Analysis, 1 fragment was expressed more abundantly in normal endometrium and 5 fragments were highly expressed in endometrial carcinoma. CONCLUSION： Combined LCM with DD-PCR, several fragments specially related to endometrial carcinoma genesis, such as CDK-7, have obtained.