Chen Xiaobo, Sun Hongwen, Zhao Yunping, Zhang Jing, Xiong Guosheng, Cui Yue, Lei Changcheng
Department of Thoracic Surgery, The First Affiliated Hospital of Kunming Medical University, No 295 Xichang Road, Kunming 650032, Yunnan, China.
Open Med (Wars). 2021 Jan 4;16(1):104-116. doi: 10.1515/med-2021-0001. eCollection 2021.
The aim of this study was to investigate the circ_0004370 expression in EC, its effects on cell proliferation, apoptosis, migration, invasion, and epithelial-mesenchymal transition (EMT) process, and the underlying regulatory mechanisms in EC.
The protein levels of COL1A1 and EMT-related proteins were detected by western blot. The role of circ_0004370 on cell viability, proliferation, and apoptosis was analyzed by Cell Counting Kit-8 (CCK-8) assay, colony formation assay, and flow cytometry, respectively. The transwell assay was used to examine cell migration and invasion. The binding sites between miR-1301-3p and circ_0004370 or COL1A1 were predicted by starbase software and confirmed by dual-luciferase reporter assay and RNA pull-down assay.
We discovered that circ_0004370 was remarkably upregulated in EC tissues and cells. Knockdown of circ_0004370 inhibited cell proliferation, migration as well as invasion, and promoted apoptosis , while its effect was rescued by miR-1301-3p inhibition. And circ_0004370 mediated the EMT process in EC cells. Moreover, we explored its regulatory mechanism and found that circ_0004370 directly bound to miR-1301-3p and COL1A1 was verified as a target of miR-1301-3p. COL1A1 was highly expressed in EC cells and upregulation of COL1A1 reversed the effects of miR-1301-3p on cell proliferation, migration, invasion, and apoptosis. In addition, silencing of circ_0004370 reduced tumor volumes and weights . We showed that circ_0004370/miR-1301-3p/COL1A1 axis played the critical role in EC to regulate the cell activities.
Circ_0004370 promotes EC proliferation, migration and invasion, and EMT process and suppresses apoptosis by regulating the miR-1301-3p/COL1A1 axis, indicating that circ_0004370 may be used as a potential therapeutic target for EC.
本研究旨在探讨circ_0004370在子宫内膜癌（EC）中的表达情况，其对细胞增殖、凋亡、迁移、侵袭以及上皮-间质转化（EMT）过程的影响，以及在EC中的潜在调控机制。
采用蛋白质印迹法检测COL1A1和EMT相关蛋白的水平。分别通过细胞计数试剂盒-8（CCK-8）检测、集落形成实验和流式细胞术分析circ_0004370对细胞活力、增殖和凋亡的作用。采用Transwell实验检测细胞迁移和侵袭能力。通过starbase软件预测miR-1301-3p与circ_0004370或COL1A1之间的结合位点，并通过双荧光素酶报告基因实验和RNA下拉实验进行验证。
我们发现circ_0004370在EC组织和细胞中显著上调。敲低circ_0004370可抑制细胞增殖、迁移和侵袭，并促进细胞凋亡，而miR-1301-3p抑制可挽救其作用。并且circ_0004370介导了EC细胞中的EMT过程。此外，我们探索了其调控机制，发现circ_0004370直接与miR-1301-3p结合，且COL1A1被证实为miR-1301-3p的靶标。COL1A1在EC细胞中高表达，COL1A1的上调逆转了miR-1301-3p对细胞增殖、迁移、侵袭和凋亡的影响。此外，沉默circ_0004370可减小肿瘤体积和重量。我们表明circ_0004370/miR-130-3p/COL1A1轴在EC中对调节细胞活性起关键作用。
Circ_0004370通过调节miR-1301-3p/COL1A1轴促进EC的增殖、迁移和侵袭以及EMT过程，并抑制细胞凋亡，表明circ_0004370可能作为EC的潜在治疗靶点。
