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目的筛选在着床窗口期子宫内膜差异表达的基因,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 技术验证其中的核糖体蛋白(RP)L7基因、RPL7假基因(RPL7p)、RPL19基因、酪氨酸3-单氧化酶/色氨酸5-单氧化酶激活蛋白 zeta 多肽(YWHAZ)基因在增生期晚期和分泌期中期子宫内膜中的表达。结果对消减文库中50个差异表达的克隆进行测序与同源性比较,筛选出35个在增生期晚期和分泌期中期有差异表达的基因。其中23个为已知功能的人类基因,12个为未知功能基因。RPL7、RPL7p、RPL19和 YWHAZ 基因在分泌期中期的表达水平有不同程度的升高,分别为0.75±0.21、1.72±0.30、1.23±0.31、1.28±0.08,分别与增生期晚期(分别为0.45±0.12、1.04±0.10、0.74±0.21、1.09±0.12)比较,差异均有统计学意义(P<0.05)。结论应用 SSH 可有效地筛选出着床窗口期子宫内膜差异表达基因,为今后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。
Objective To screen the differentially expressed genes of endometrium in the implantation window to understand the molecular basis of the formation of endometrial receptivity. Methods Suppression subtractive hybridization (SSH) was used to establish a subtractive library of endometrial implantation window genes, and the differentially expressed genes were identified by sequencing and homology. The ribosomes were identified by RT-PCR (RP) L7 gene, RPL7 pseudogene (RPL7p), RPL19 gene, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activator protein zeta polypeptide (YWHAZ) gene in the late proliferative phase and the secretory phase Endometrial expression. Results Fifty differentially expressed clones in subtracted libraries were sequenced and homologous compared to 35 differentially expressed genes in late proliferative phase and secretory phase. 23 of them are known human genes and 12 are unknown genes. The expression levels of RPL7, RPL7p, RPL19 and YWHAZ in the middle of secretory phase were increased to some extent, which were 0.75 ± 0.21,1.72 ± 0.30,1.23 ± 0.31,1.28 ± 0.08 respectively, which were respectively associated with the late proliferative phase (0.45 ± 0.12,1.04 ± 0.10,0.74 ± 0.21,1.09 ± 0.12), the differences were statistically significant (P <0.05). Conclusion SSH can effectively screen out endometrial differentially expressed genes at the implantation window and provide new clues for further research on the molecular mechanism of endometrial receptivity.