Department of Endocrinology, Shanxi Bethune Hospital, Taiyuan City, Shanxi Province, 030032, People's Republic of China.
Department of Endocrinology, Huhhot First Hospital, Huhhot, The Inner Mongolia, Autonomous Region, 010020, People's Republic of China.
J Ovarian Res. 2021 Nov 5;14(1):151. doi: 10.1186/s13048-021-00905-x.
miR-146b-5p has been reported to participate in premature ovarian failure (POF) in mice. However, its role in POF patients is unclear. We predicted that miR-146b-5p might interact with lncRNA DLEU1, a crucial player in ovarian cancer. We then explored the interaction between DLEU1 and miR-146b-5p.
Expression of DLEU1 and miR-146b-5p in POF and control ovary tissues was determined by RT-qPCR. The subcellular location of DLEU1 in human KGN cells was analyzed using subcellular fractionation assays. The direct interaction between DLEU1 and miR-146b-5p was analyzed using RNA pull-down assays. The role of DLEU1 in miR-146a expression was analyzed using overexpression assay. Cell proliferation was analyzed using cell apoptosis assay.
Increased DLEU1 expression and decreased miR-146b-5p expression were observed in POF. DLEU1 directly interacted with MiR-146b-5p and was expressed in both nuclear and cytoplasm samples of KGN cells. In KGN cells, DLEU1 and miR-146b-5p failed to regulate the expression of each other. However, DLEU1 promoted cell apoptosis and reduced the inhibitory effects of miR-146b-5p on cell apoptosis.
DLEU1 is overexpressed in POF and sponges miR-146b-5p to increase KGN cell apoptosis.
已有研究表明 miR-146b-5p 参与了小鼠的卵巢早衰（POF）。然而，其在 POF 患者中的作用尚不清楚。我们推测 miR-146b-5p 可能与 lncRNA DLEU1 相互作用，DLEU1 是卵巢癌中的关键调控因子。因此，我们进一步探索了 DLEU1 与 miR-146b-5p 之间的相互作用。
采用 RT-qPCR 检测 POF 和对照卵巢组织中 DLEU1 和 miR-146b-5p 的表达。采用亚细胞分离实验分析 DLEU1 在人 KGN 细胞中的亚细胞定位。采用 RNA 下拉实验分析 DLEU1 与 miR-146b-5p 的直接相互作用。采用过表达实验分析 DLEU1 对 miR-146a 表达的影响。采用细胞凋亡实验分析细胞增殖情况。
POF 患者中 DLEU1 表达增加，miR-146b-5p 表达降低。DLEU1 与 miR-146b-5p 直接相互作用，并且在 KGN 细胞的核和细胞质样本中均有表达。在 KGN 细胞中，DLEU1 和 miR-146b-5p 不能相互调节对方的表达。然而，DLEU1 促进细胞凋亡，并降低了 miR-146b-5p 对细胞凋亡的抑制作用。
DLEU1 在 POF 中过表达，通过海绵吸附 miR-146b-5p 增加 KGN 细胞凋亡。
