子宫内膜异位症相关性不孕患者的子宫内膜差异表达基因cDNA文库的构建和筛选

目的:筛选子宫内膜异位症相关性不孕患者在着床窗口期子宫内膜差异表达基因,以了解子宫内膜容受性形成的分子基础。方法:采用抑制性消减杂交技术(SSH),建立人子宫内膜着床窗口期基因表达的消减文库,并通过测序和同源性比较确定差异表达的基因;用RT-PCR技术验证其中的翻译控制肿瘤蛋白(TCTP1)基因、载脂蛋白D(ApoD)基因、促红细胞生成素(EPO)基因在分泌期中期子宫内膜中的表达。结果:对消减文库中20个差异表达的克隆进行测序与同源性比较,筛选出8个在分泌期中期有差异表达的基因。其中TCTP1、ApoD、EPO基因在分泌期中期的表达水平有不同程度的升高,分别为1.35±0.12、1.24±0.10、1.05±0.23,分别与正常对照组的1.05±0.23、0.75±0.16、0.68±0.08比较,差异均有统计学意义(P<0.05)。结论:应用SSH可有效地筛选出内异症相关性不孕患者着床窗口期子宫内膜差异表达基因,为进一步研究子宫内膜容受性形成的分子机理提供新的线索。

Construction and screening of cDNA library of differentially expressed genes of endometrium in endometriosis-associated infertility

Objective:To identify the genes differentially expressed in the window of implantation and explore the molecular basis of the development of endometrial receptivity in infertile patients with endometriosis.Methods:A subtracted cDNA library in window of implantation was constructed by suppression subtractive hybridization(SSH) method,then differentially expressed genes of endometrium were identified by gene sequencing and homology comparison.The differential expression of translationally controlled tumor protein(TCTP1) gene,apolipoprotein D(ApoD) gene and erythropoietin(EPO) gene were further confirmed by RT-PCR.Results:After gene sequencing and homology comparison of 20 clones,8 differentially expressed genes were detected in window of implantation.The expression levels of TCTP1,ApoD and EPO in window of implantation were 1.35±0.12,1.24±0.10 and 1.05±0.23,respectively,which were significantly higher than those in control group(1.05±0.23,0.75±0.16,0.68±0.08)(P<0.05).Conclusion:SSH is a useful technique to detect differential expression genes of endometrium in window of implantation in endometriosis-associated infertility,which provides a new clue to investigate the molecular basis of the development of endometrial receptivity.