Feng Fen, Wang Jing, Bao Riqiang, Li Long, Tong Xiating, Han Suo, Zhang Hongdan, Wen Weihui, Xiao Li, Zhang Chunping
Department of Cell Biology, College of Medicine, Nanchang University, Nanchang, China.
Department of Microbiology, College of Medicine, Nanchang University, Nanchang, China.
Front Cell Dev Biol. 2020 Jun 30;8:481. doi: 10.3389/fcell.2020.00481. eCollection 2020.
LncPrep + 96kb is a novel long non-coding RNA expressed in murine granulosa cells with two transcripts that are 2.2 and 2.8 kb in length. However, the potential roles of lncPrep + 96kb in granulosa cells remain poorly understood. In this study, we investigated the effect of the lncPrep + 96kb 2.2 kb transcript on granulosa cells through the overexpression and knockdown of lncPrep + 96kb 2.2 kb. We found that lncPrep + 96kb 2.2 kb inhibited aromatase expression and estradiol production. Endothelial differentiation-related factor 1 (EDF1) is an evolutionarily conserved transcriptional coactivator. We found that EDF1 knockdown inhibited aromatase expression and estradiol production. The RNA immunoprecipitation results also showed that lncPrep + 96kb 2.2 kb can bind to EDF1 and that overexpression of lncPrep + 96kb 2.2 kb induced the translocation of EDF1 from the nucleus to the cytoplasm. The CatRAPID signature revealed that the 1,979-2,077 and 603-690 nucleotide positions in lncPrep + 96kb 2.2 kb were potential binding sites for EDF1. We found that mutating the 1,979-2,077 site rescued the effects of lncPrep + 96kb 2.2 kb on aromatase expression and estradiol production. In conclusion, we are the first to report that specific expression of lncPrep + 96kb 2.2 kb in granulosa cells inhibits the production of estradiol by influencing the localization of EDF1 in granulosa cells. The 1,979-2,077 site of lncPrep + 96kb 2.2 kb contributes to the ability to bind to EDF1.
LncPrep + 96kb是一种在小鼠颗粒细胞中表达的新型长链非编码RNA，有两种转录本，长度分别为2.2 kb和2.8 kb。然而，lncPrep + 96kb在颗粒细胞中的潜在作用仍知之甚少。在本研究中，我们通过lncPrep + 96kb 2.2 kb的过表达和敲低来研究其对颗粒细胞的影响。我们发现lncPrep + 96kb 2.2 kb抑制芳香化酶表达和雌二醇生成。内皮分化相关因子1（EDF1）是一种进化上保守的转录共激活因子。我们发现敲低EDF1可抑制芳香化酶表达和雌二醇生成。RNA免疫沉淀结果还显示，lncPrep + 96kb 2.2 kb可与EDF1结合，且lncPrep + 96kb 2.2 kb的过表达诱导EDF1从细胞核转移至细胞质。CatRAPID特征显示，lncPrep + 96kb 2.2 kb中的1979 - 2077和603 - 690核苷酸位置是EDF1的潜在结合位点。我们发现突变1979 - 2077位点可挽救lncPrep + 96kb 2.2 kb对芳香化酶表达和雌二醇生成的影响。总之，我们首次报道lncPrep + 96kb 2.2 kb在颗粒细胞中的特异性表达通过影响EDF1在颗粒细胞中的定位来抑制雌二醇生成。lncPrep + 96kb 2.2 kb的1979 - 2077位点有助于其与EDF1结合的能力。
