Expression Analysis of Long Noncoding RNA in the Cumulus Granulosa Cells of PCOS Based on Transcriptome Sequencing

Abstract

Objective: The present study aims to provide new ideas for research on the molecular mechanism of polycystic ovary syndrome (PCOS) by (1) investigating the expression characteristics of long non-coding RNAs (lncRNAs) in cumulus granulosa cells (GCs) in patients with PCOS and (2) conducting in vitro validation. Methods: The differential expression profiles of lncRNAs in GCs from patients with PCOS and women with normal ovulation were obtained by sequencing using the HiSeq platform. The key lncRNAs were screened, and the target genes were verified by real-time polymerase chain reaction (RT-PCR) and Western blot in GCs of different quality (high-quality embryos and non-high-quality embryos). Results: A total of 204 significantly differentially expressed lncRNAs were identified using the results of transcriptome gene sequencing;The expression level was up-regulated in 83 and down-regulated in 121. The RT-PCR validation showed that the expressions of target lncRNA ENST00000433673 and messenger RNA (mRNA) NPY1R (Neuropeptide Y Receptor Y1) were significantly higher in the GCsof non-high-quality embryos from patients with PCOS than in the GCs of non-high-quality embryos from healthy subjects;Meanwhile, the expression of the NPY1R protein was significantly higher in the GCs of non-high-quality embryos from healthy subjects than in the GCs of non-high-quality embryos from patients with PCOS. For high-quality embryos, the expressions of lncRNA ENST00000433673 and mRNA NPY1R did not change significantly in GCs from both healthy subjects and patients with PCOS. The expression of NPY1R protein have similar pattern with NPY1R mRNA in different groups, the expression level of NPY1R protein was the highest in control group, and the expression of lncRNA ENST00000433673 increased while the protein expression of NPY1R decreased. Conclusions: lncRNA ENST00000433673 might have important regulatory roles during processes such as embryonic development;This suggests that lncRNAs might be involved in the pathogenesis of PCOS.