First School of Clinical Medicine, Shandong University of Traditional Chinese Medicine, Jinan, 250011, China.
Department of Obstetrics and Gynaecology, BC Children's Hospital Research Institute, University of British Columbia, Vancouver, British Columbia, Canada.
J Ovarian Res. 2022 Oct 29;15(1):119. doi: 10.1186/s13048-022-01053-6.
Infertility is a global reproductive-health problem, and diminished ovarian reserve (DOR) is one of the common causes of female infertility. Long noncoding RNAs (lncRNAs) are crucial regulators of numerous physiological and pathological processes in humans. However, whether lncRNAs are involved in the development of DOR remains to be elucidated.
Ovarian granulosa cells (OGCs) extracted from infertile women with DOR and from women with normal ovarian reserve (NOR) were subjected to high-throughput sequencing. Comprehensive bioinformatics analysis was conducted to identify the differential expression of messenger RNAs (mRNAs) and lncRNAs. Sequencing results were validated by the selection of lncRNAs and mRNAs using real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR).
Compared with the NOR group, a total of 244 lncRNAs were upregulated (53 known and 191 novel), and 222 lncRNAs were downregulated (36 known and 186 novel) in the DOR group. Similarly, 457 mRNAs had differential expression between the two groups. Of these, 169 were upregulated and 288 were downregulated. Bioinformatics analysis revealed that the differentially expressed genes of mRNA and lncRNAs were considerably enriched in "cell adhesion and apoptosis", "steroid biosynthesis", and "immune system". A co-expression network comprising lncRNAs and their predicted target genes revealed the possible involvement of the "thyroid hormone signaling pathway" and "protein binding, digestion and absorption" in DOR pathogenesis. The expression of SLC16A10 was positively regulated by multiple lncRNAs. After RT-qPCR validation of seven differentially expressed lncRNAs and mRNAs, respectively, the expression of lncRNA NEAT1, GNG12, ZEB2-AS1, and mRNA FN1, HAS3, RGS4, SUOX were in accordance with RNA-sequencing.
We presented the first data showing that the expression profiles of lncRNA and mRNA in OGCs between NOR and DOR patients using RNA sequencing. The lncRNAs and mRNAs that we identified may serve as novel diagnostic biomarkers for patients with DOR.
不孕是一个全球性的生殖健康问题，而卵巢储备功能降低（DOR）是女性不孕的常见原因之一。长链非编码 RNA（lncRNA）是人类许多生理和病理过程的关键调节因子。然而，lncRNA 是否参与 DOR 的发生仍有待阐明。
从患有 DOR 的不孕女性和具有正常卵巢储备（NOR）的女性中提取卵巢颗粒细胞（OGC），并进行高通量测序。采用综合生物信息学分析方法，鉴定信使 RNA（mRNA）和 lncRNA 的差异表达。通过实时逆转录定量聚合酶链反应（RT-qPCR）选择 lncRNA 和 mRNA 对测序结果进行验证。
与 NOR 组相比，DOR 组共有 244 个 lncRNA 上调（53 个已知和 191 个新的），222 个 lncRNA 下调（36 个已知和 186 个新的）。同样，两组之间有 457 个 mRNA 存在差异表达。其中，169 个上调，288 个下调。生物信息学分析表明，mRNA 和 lncRNA 的差异表达基因在“细胞黏附和凋亡”、“类固醇生物合成”和“免疫系统”中显著富集。包含 lncRNA 及其预测靶基因的共表达网络显示，“甲状腺激素信号通路”和“蛋白质结合、消化和吸收”可能参与 DOR 的发病机制。SLC16A10 的表达受多个 lncRNA 的正向调节。通过分别对 7 个差异表达的 lncRNA 和 mRNA 进行 RT-qPCR 验证，lncRNA NEAT1、GNG12、ZEB2-AS1 和 mRNA FN1、HAS3、RGS4、SUOX 的表达与 RNA 测序结果一致。
我们首次使用 RNA 测序展示了 NOR 和 DOR 患者 OGC 中 lncRNA 和 mRNA 表达谱。我们鉴定的 lncRNA 和 mRNA 可能作为 DOR 患者的新型诊断生物标志物。
