Li Yanfang, Wu Zhiqing, Shen Jianlin
Department of Pediatric Surgery, Affiliated Hospital of Putian University, #181 Meiyuan East Road, Putian, 351100, Fujian, China.
Department of Orthopedics, Affiliated Hospital of Putian University, #999 Dongzhen East Road, Putian, 351100, Fujian, China.
Regen Ther. 2022 Jun 22;21:114-121. doi: 10.1016/j.reth.2022.06.004. eCollection 2022 Dec.
Accumulating evidence has shown that circular RNAs (circRNAs) have indispensable functions during tumor progression by regulating gene expression. A previous study found that upregulation of hsa_circ_0000885 indicated a poor clinical outcome of osteosarcoma (OS). However, the regulatory mechanism of this process is unclear.
This investigation aimed to elucidate how hsa_circ_0000885 regulated OSs. The study used RT-qPCR to investigate hsa_circ_0000885 expression in OS cells. We conducted luciferase reporter assays and analyses to confirm the hsa_circ_0000885 downstream target. We transfected OS cells using different vectors and used Transwell migration, colony formation, western blotting, Matrigel invasion, proliferation, tumorigenesis, and metastasis assays to identify the role of hsa_circ_0000885 in OS.
The results showed that hsa_circ_0000885 expression altered OS cell lines, and that hsa_circ_0000885 downregulation suppressed OS cell proliferation and invasion using and experiments. Luciferase reporter assays verified that miR-16-5p and E2F3 were downstream targets of hsa_circ_0000885. E2F3 overexpression or miR-16-5p inhibition reversed OS cell invasion and proliferation after silencing hsa_circ_0000885. Furthermore, hsa_circ_0000885 affected cancer stem cell differentiation by regulating miR-16-5p/E2F3.
Overall, the results showed that hsa_circ_0000885 downregulation suppressed OS progression and metastasis via regulating E2F3 expression and sponging miR-16-5p.
越来越多的证据表明,环状RNA(circRNA)通过调节基因表达在肿瘤进展过程中发挥着不可或缺的作用。先前的一项研究发现,hsa_circ_0000885的上调表明骨肉瘤(OS)的临床预后较差。然而,这一过程的调控机制尚不清楚。
本研究旨在阐明hsa_circ_0000885如何调节骨肉瘤。该研究使用逆转录定量聚合酶链反应(RT-qPCR)来检测骨肉瘤细胞中hsa_circ_0000885的表达。我们进行了荧光素酶报告基因检测和分析,以确认hsa_circ_0000885的下游靶点。我们使用不同的载体转染骨肉瘤细胞,并通过Transwell迁移、集落形成、蛋白质免疫印迹法、基质胶侵袭、增殖、肿瘤发生和转移检测,来确定hsa_circ_0000885在骨肉瘤中的作用。
结果表明,hsa_circ_0000885的表达改变了骨肉瘤细胞系,并且通过[具体实验名称1]和[具体实验名称2]实验发现,hsa_circ_0000885的下调抑制了骨肉瘤细胞的增殖和侵袭。荧光素酶报告基因检测证实,miR-16-5p和E2F3是hsa_circ_00