Department of Orthopaedics, First Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong, China.
Center for Precision Health, School of Medical and Health Science, Edith Cowan University, Perth, Western Australia, Australia.
J Cell Mol Med. 2024 Apr;28(8):e18269. doi: 10.1111/jcmm.18269.
Circular RNAs (circRNAs) play an important role in the progression of osteosarcoma. However, the precise function of circPVT1 in osteosarcoma remains elusive. This study aims to explore the molecular mechanism underlying the involvement of circPVT1 in osteosarcoma cells. We quantified circPVT1 expression using qRT-PCR in both control and osteosarcoma cell lines. To investigate the roles of circPVT1, miR-490-5p and HAVCR2 in vitro, we separately conducted overexpression and inhibition experiments for circPVT1, miR-490-5p and HAVCR2 in HOS and U2OS cells. Cell migration was assessed through wound healing and transwell migration assays, and invasion was measured via the Matrigel invasion assay. To elucidate the regulatory mechanism of circPVT1 in osteosarcoma, a comprehensive approach was employed, including fluorescence in situ hybridization, qRT-PCR, Western blot, bioinformatics, dual-luciferase reporter assay and rescue assay. CircPVT1 expression in osteosarcoma cell lines surpassed that in control cells. The depletion of circPVT1 resulted in a notable reduction in the in vitro migration and invasion of osteosarcoma cells. Mechanism experiments revealed that circPVT1 functioned as a miR-490-5p sequester, and directly targeted HAVCR2. Overexpression of miR-490-5p led to a significant attenuation of migration and invasion of osteosarcoma cells, whereas HAVCR2 overexpression had the opposite effect, promoting these abilities. Additionally, circPVT1 upregulated HAVCR2 expression via sequestering miR-490-5p, thereby orchestrating the migration and invasion in osteosarcoma cells. CircPVT1 orchestrates osteosarcoma migration and invasion by regulating the miR-490-5p/HAVCR2 axis, underscoring its potential as a promising therapeutic target for osteosarcoma.
环状 RNA(circRNAs)在骨肉瘤的进展中发挥着重要作用。然而,circPVT1 在骨肉瘤中的确切功能仍不清楚。本研究旨在探讨 circPVT1 参与骨肉瘤细胞的分子机制。我们使用 qRT-PCR 定量检测了对照和骨肉瘤细胞系中 circPVT1 的表达。为了研究 circPVT1、miR-490-5p 和 HAVCR2 在体外的作用,我们分别在 HOS 和 U2OS 细胞中进行了 circPVT1、miR-490-5p 和 HAVCR2 的过表达和抑制实验。通过划痕愈合和 Transwell 迁移实验评估细胞迁移,通过 Matrigel 侵袭实验测量侵袭。为了阐明 circPVT1 在骨肉瘤中的调控机制,我们采用了包括荧光原位杂交、qRT-PCR、Western blot、生物信息学、双荧光素酶报告基因检测和挽救实验在内的综合方法。骨肉瘤细胞系中 circPVT1 的表达高于对照细胞。circPVT1 的耗竭导致骨肉瘤细胞体外迁移和侵袭明显减少。机制实验表明,circPVT1 作为 miR-490-5p 的海绵发挥作用,并直接靶向 HAVCR2。过表达 miR-490-5p 导致骨肉瘤细胞迁移和侵袭能力显著减弱,而过表达 HAVCR2 则产生相反的效果,促进了这些能力。此外,circPVT1 通过海绵吸附 miR-490-5p 上调 HAVCR2 表达,从而协调骨肉瘤细胞的迁移和侵袭。circPVT1 通过调节 miR-490-5p/HAVCR2 轴来协调骨肉瘤的迁移和侵袭,这表明其作为骨肉瘤有前途的治疗靶点的潜力。