背景:比较多囊卵巢综合征(PCOS)患者和正常排卵患者接受体外受精-胚胎移植(IVF-ET)前子宫内膜中长非编码RNA(lncRNA)和信使RNA(mRNA)的表达水平。
方法:采用子宫内膜真空刮匙在接受前阶段(取卵后3天)收集PCOS组和对照组的子宫内膜组织。通过RNA测序和比对鉴定子宫内膜的LncRNA和mRNA。通过定量逆转录聚合酶链反应(qRT-PCR)在22名PCOS患者和18名排卵患者中验证了9个差异表达lncRNAs和11个mRNAs的子集。使用基因本体论(GO)和京都基因和基因组百科全书(KEGG)探索了具有差异表达模式的mRNA的功能。
结果:我们发现了687个上调的mRNA和680个下调的mRNA,与正常排卵患者相比,PCOS患者中345个上调lncRNAs和63个下调lncRNAs。qRT-PCR检测11种mRNA的表达,并验证了CXCR4、RABL6、OPN3、SYBU、IDH1、NOP10在PCOS患者中显著升高,ZEB1的表达明显下降。进行qRT-PCR以检测9种lncRNAs的表达,并验证了这7种lncRNAsIDH1-AS1,PCAT14,FTX的表达,DANCR,PRKCQ-AS1、SNHG8、TPT1-AS1在PCOS患者中显著增强。生物信息学分析显示,差异表达基因(DEGs)涉及KEGG通路,PI3K-Akt通路,代谢途径,Jak-STAT通路,丙酮酸代谢,内质网中的蛋白质加工,氧化磷酸化和蛋白酶体。GO分级上调参与ATP代谢过程,氧化磷酸化,RNA分解代谢过程,GO分类的下调是对皮质类固醇的反应,类固醇激素,和T细胞激活。
结论:我们的结果确定了PCOS患者在接受前阶段子宫内膜lncRNAs和mRNAs的特征和表达谱,也就是卵母细胞取出后的第3天。发现子宫内膜容受性障碍的可能通路及相关基因,这些lncRNAs可能在接受前阶段被开发为子宫内膜的预测生物标志物。
BACKGROUND: To compare the expression levels of long non-coding RNA (lncRNA) and messenger RNA (mRNA) in pre-receptive endometrium between patients with Polycystic Ovary Syndrome (PCOS)and normal ovulation undergoing in vitro fertilization-embryo transfer (IVF-ET).
METHODS: Endometrial tissues were collected with endometrial vacuum curette in pre-receptive phase (3 days after oocytes retrieval) from PCOS and control groups. LncRNAs and mRNAs of endometrium were identified via RNA sequencing and alignments. A subset of 9 differentially expressed lncRNAs and 11 mRNAs were validated by quantitative reverse transcription polymerase chain reaction(qRT-PCR)in 22 PCOS patients and 18 ovulation patients. The function of mRNAs with differential expression patterns were explored using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG).
RESULTS: We found out 687 up-regulated and 680 down-regulated mRNAs, as well as 345 up-regulated and 63 down-regulated lncRNAs in the PCOS patients in contrast to normal ovulation patients. qRT-PCR was used to detect the expression of 11 mRNAs, and validated that the expression of these 6 mRNAs CXCR4, RABL6, OPN3, SYBU, IDH1, NOP10 were significantly elevated among PCOS patients, and the expression of ZEB1 was significantly decreased. qRT-PCR was performed to detect the expression of 9 lncRNAs, and validated that the expression of these 7 lncRNAs IDH1-AS1, PCAT14, FTX, DANCR, PRKCQ-AS1, SNHG8, TPT1-AS1 were significantly enhanced among PCOS patients. Bioinformatics analysis showed that differentially expressed genes (DEGs) involved KEGG pathway were tyrosine metabolism, PI3K-Akt pathway, metabolic pathway, Jak-STAT pathway, pyruvate metabolism, protein processing in endoplasmic reticulum, oxidative phosphorylation and proteasome. The up-regulation of GO classification was involved in ATP metabolic process, oxidative phosphorylation, RNA catabolic process, and down-regulation of GO classification was response to corticosteroid, steroid hormone, and T cell activation.
CONCLUSIONS: Our results determined the characteristics and expression profile of endometrial lncRNAs and mRNAs in PCOS patients in pre-receptive phase, which is the day 3 after oocytes retrival. The possible pathways and related genes of endometrial receptivity disorders were found, and those lncRNAs may be developed as a predictive biomarker of endometrium in pre-receptive phase.