Li Xiaoli, Lu Shenyi, Jiang Yan, Xu Kai, He Jingzhi, Qiu Zhangli, Ying Pian
Hangzhou Suntaihe Traditional Chinese Medicine and Medicine Museum, Hangzhou, China.
Department of Gynecology and Obstetrics, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, China.
Gynecol Obstet Invest. 2022;87(5):286-298. doi: 10.1159/000525889. Epub 2022 Aug 10.
The aim of our study was to explore the role of circular RNA_0061140 (circ_0061140) in adenomyosis progression and its associated mechanism.
We first analyzed the expression pattern of circ_0061140 in endometrial tissues of adenomyosis patients (n = 27) and uterine fibroid patients (n = 15). Loss-of-function experiments were conducted to analyze the biological roles of circ_0061140 in regulating the viability, apoptosis, proliferation, migration, and invasion of endometrial epithelial cells. The downstream microRNA (miRNA)/messenger RNA (mRNA) axis of circ_0061140 was predicted by bioinformatics tool Starbase, and its working mechanism was verified by rescue experiments.
Cell viability, apoptosis, proliferation, invasion, and migration were assessed by cell counting kit-8 assay, flow cytometry analysis, 5-ethynyl-2'-deoxyuridine assay, transwell assay, and scratch test. The binding relationship between miR-141-3p and circ_0061140 or lin-28 homolog B (LIN28B) was verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay.
Circ_0061140 expression was upregulated in adenomyosis patients. Circ_0061140 knockdown suppressed the viability, proliferation, invasion, and migration and triggered the apoptosis of endometrial epithelial cells. Circ_0061140 served as a miRNA sponge for miR-141-3p, and miR-141-3p silencing partly reversed circ_0061140 knockdown-induced effects in endometrial epithelial cells. miR-141-3p directly interacted with LIN28B mRNA. LIN28B overexpression partly diminished miR-141-3p overexpression-mediated influences in endometrial epithelial cells. Circ_0061140 knockdown downregulated LIN28B expression by elevating miR-141-3p level in endometrial epithelial cells.
The functional verification of circ_0061140/miR-141-3p/LIN28B axis was merely conducted in vitro.
Circ_0061140 contributed to adenomyosis progression by binding to miR-141-3p to induce LIN28B expression in vitro.
本研究旨在探讨环状RNA_0061140（circ_0061140）在子宫腺肌病进展中的作用及其相关机制。
我们首先分析了子宫腺肌病患者（n = 27）和子宫肌瘤患者（n = 15）子宫内膜组织中circ_0061140的表达模式。进行功能丧失实验以分析circ_0061140在调节子宫内膜上皮细胞活力、凋亡、增殖、迁移和侵袭中的生物学作用。通过生物信息学工具Starbase预测circ_0061140的下游微小RNA（miRNA）/信使核糖核酸（mRNA）轴，并通过挽救实验验证其作用机制。
通过细胞计数试剂盒-8检测、流式细胞术分析、5-乙炔基-2'-脱氧尿苷检测、Transwell检测和划痕试验评估细胞活力、凋亡、增殖、侵袭和迁移。通过双荧光素酶报告基因检测和RNA免疫沉淀（RIP）检测验证miR-141-3p与circ_0061140或线性28同源物B（LIN28B）之间的结合关系。
子宫腺肌病患者中circ_0061140表达上调。circ_0061140敲低抑制了子宫内膜上皮细胞的活力、增殖、侵袭和迁移，并引发了细胞凋亡。circ_0061140作为miR-141-3p的miRNA海绵，miR-141-3p沉默部分逆转了circ_0061140敲低对子宫内膜上皮细胞的影响。miR-141-3p直接与LIN28B mRNA相互作用。LIN28B过表达部分减弱了miR-141-3p过表达对子宫内膜上皮细胞的影响。circ_0061140敲低通过提高子宫内膜上皮细胞中miR-141-3p水平下调LIN28B表达。
circ_0061140/miR-141-3p/LIN28B轴的功能验证仅在体外进行。
在体外，circ_0061140通过与miR-141-3p结合诱导LIN28B表达，促进子宫腺肌病进展。
