Gao Chunsheng, Wang Xiaowei, Yan Huichao, Zeng Ge, Chen Yan, Gao Jun
Department of Orthopaedics, The Third People's Hospital of Hubei Province, Wuhan, China.
DNA Cell Biol. 2025;44(3):153-160. doi: 10.1089/dna.2024.0245. Epub 2025 Jan 8.
Exosome-delivered circular RNAs (circRNAs) are recognized as a key mechanism that regulates osteosarcoma (OS) progression. The purpose of this study is to discover the role of a novel circRNA hsa_circ_0000116 from exosomes in OS progression. Transmission electron microscopy, nanoparticle tracking analysis, and western blotting were used to identify the exosomes isolated from two OS cell lines (HOS and MG-63). After coculturing exosomes with OS cells and transfecting hsa_circ_0000116 knockdown vector into OS cells, cell function experiments, including cell counting kit-8, wound healing, and Transwell experiments, were performed to assess the change of OS cell malignant phenotype. In addition, the levels of PI3K/Akt/mTOR and p38/MAPK pathways-associated proteins were measured using western blotting. Exosomes with around 100 nm in diameter were successfully isolated from HOS and MG-63 cells, and promote OS cells to proliferate, migrate, and invade. hsa_circ_0000116 was upregulated in OS-derived exosomes, and silencing hsa_circ_0000116 declined the exosome-induced OS cell malignancy. In addition, inhibiting hsa_circ_0000116 effectively inhibited exosome-mediated activation of PI3K/Akt/mTOR and p38/MAPK pathways. In conclusion, exosomal hsa_circ_0000116 can facilitate OS cell malignancy by inducing the activation of PI3K/Akt/mTOR and p38/MAPK pathways. The findings of this study may identify novel molecular mechanisms driving OS progression and provide novel therapeutic targets for OS.
外泌体传递的环状RNA（circRNAs）被认为是调节骨肉瘤（OS）进展的关键机制。本研究的目的是发现外泌体中一种新型circRNA hsa_circ_0000116在OS进展中的作用。采用透射电子显微镜、纳米颗粒跟踪分析和蛋白质免疫印迹法鉴定从两种OS细胞系（HOS和MG-63）中分离出的外泌体。将外泌体与OS细胞共培养，并将hsa_circ_0000116敲低载体转染到OS细胞中后，进行细胞功能实验，包括细胞计数试剂盒-8、伤口愈合和Transwell实验，以评估OS细胞恶性表型的变化。此外，使用蛋白质免疫印迹法检测PI3K/Akt/mTOR和p38/MAPK通路相关蛋白的水平。成功从HOS和MG-63细胞中分离出直径约100 nm的外泌体，其可促进OS细胞增殖、迁移和侵袭。hsa_circ_0000116在OS来源的外泌体中上调，沉默hsa_circ_0000116可降低外泌体诱导的OS细胞恶性程度。此外，抑制hsa_circ_0000116可有效抑制外泌体介导的PI3K/Akt/mTOR和p38/MAPK通路的激活。总之，外泌体hsa_circ_0000116可通过诱导PI3K/Akt/mTOR和p38/MAPK通路的激活促进OS细胞恶性程度。本研究结果可能确定驱动OS进展的新分子机制，并为OS提供新的治疗靶点。
