Department of Gynecology, the First Hospital of Changsha, Changsha, Hunan 410005, PR China.
Department of Intensive Care, the First Hospital of Changsha, Changsha, Hunan 410005, PR China.
Reprod Biol. 2022 Jun;22(2):100643. doi: 10.1016/j.repbio.2022.100643. Epub 2022 Apr 30.
Circular RNA ATRNL1 (circATRNL1) has been implicated in epithelial-mesenchymal transition (EMT) during endometriosis. Given the existing literature and our predictions through starBase in this research, it was assumed that circATRNL1 might orchestrate the microRNA (miR)- 103a-3p/acid-sensing ion channel 1 (ASIC1) axis to control EMT in endometriosis. To verify our hypothesis, we detect circATRNL1, miR-103a-3p, and ASIC1 expression in endometrial cancer cells (HEC-B, AN3-CA, KLE, HEC1-A, and Ishikawa). Ishikawa cells with the highest circATRNL1 level were selected as subjects, where circATRNL1, miR-103a-3p, or ASIC1 expression was knocked down. Scratch and Transwell assays were applied to assess cell migration and invasion, and CCK-8 and colony formation assays to detect cell proliferation. Western blot was used to measure E-cadherin, N-cadherin, Vimentin, and Slug expression to evaluate the EMT state. Furthermore, the binding of miR-103a-3p to circATRNL1 or ASIC1 was validated by luciferase reporter assay. CircATRNL1 and ASIC1 were upregulated but miR-103a-3p was downregulated in endometrial cancer cells. Mechanistically, circATRNL1 bound to miR-103a-3p to upregulate a target gene of miR-103a-3p, ASIC1. CircATRNL1 silencing contributed to the decline of proliferation, invasion, migration, and EMT in Ishikawa cells, while miR-103a-3p inhibitor reversed those changes. In addition, the EMT process was aggravated when miR-103a-3p was inhibited and this process was suppressed by silencing ASIC1 in the presence of downregulated miR-101a-3p. Our study supported that circATRNL1 might be a novel therapeutic candidate target for endometriosis treatment and provided unique insights into the molecular basis concerning the pathogenesis of endometriosis.
环状 RNA ATRNL1（circATRNL1）已被牵连在内异症的上皮间质转化（EMT）过程中。鉴于现有文献和我们在本研究中通过 starBase 的预测，假设 circATRNL1 可能通过调控 microRNA（miR）-103a-3p/酸敏离子通道 1（ASIC1）轴来控制内异症的 EMT。为了验证我们的假说，我们检测了子宫内膜癌细胞（HEC-B、AN3-CA、KLE、HEC1-A 和 Ishikawa）中 circATRNL1、miR-103a-3p 和 ASIC1 的表达。选择具有最高 circATRNL1 水平的 Ishikawa 细胞作为研究对象，敲低 circATRNL1、miR-103a-3p 或 ASIC1 的表达。应用划痕和 Transwell 实验评估细胞迁移和侵袭，CCK-8 和集落形成实验检测细胞增殖。Western blot 用于测量 E-钙黏蛋白、N-钙黏蛋白、波形蛋白和 Slug 的表达，以评估 EMT 状态。此外，通过荧光素酶报告基因实验验证了 miR-103a-3p 与 circATRNL1 或 ASIC1 的结合。在子宫内膜癌细胞中，circATRNL1 和 ASIC1 上调，但 miR-103a-3p 下调。在机制上，circATRNL1 与 miR-103a-3p 结合，上调 miR-103a-3p 的靶基因 ASIC1。circATRNL1 沉默促进了 Ishikawa 细胞增殖、侵袭、迁移和 EMT 的下降，而 miR-103a-3p 抑制剂逆转了这些变化。此外，当抑制 miR-103a-3p 时，EMT 过程加剧，而在下调的 miR-103a-3p 存在下沉默 ASIC1 则抑制了这一过程。我们的研究支持 circATRNL1 可能成为内异症治疗的一种新的治疗靶点，并为内异症发病机制的分子基础提供了独特的见解。
