Department of Orthopedics, Zhengzhou University People's Hospital, Henan Provincial People's Hospital.
Department of Pathology, Zhengzhou University People's Hospital, Henan Provincial People's Hospital.
Biol Pharm Bull. 2022;45(9):1283-1290. doi: 10.1248/bpb.b22-00198.
Ubiquitin-specific peptidase 9X (USP9X) has been reported to be closely associated with the formation and progression of a variety of malignant tumors. However, the mechanism by which USP9X is involved in osteosarcoma and development has not been clearly studied. This work aimed to probe the influence of USP9X on osteosarcoma cell proliferation, migration and invasion. This study recruited sixty-seven patients with histologically definited osteosarcoma. Osteosarcoma samples and cell-line were used to reflect the expression level of USP9X. Analysis of cell proliferation by thiazolium blue (MTT) assays. Transwell experiments and wound healing were used to verify cell migration and invasion capabilities. The effect of USP9X was investigated through in vivo experiments. USP9X-related pathway proteins were detected by Western blot and quantitative real-time PCR (qRT-PCR). The expression of USP9X in osteosarcoma was higher than that in adjacent tissues. The overall survival of patients with USP9X-negative patients was better than that of patients with USP9X-positive. The growth of osteosarcoma cells in vivo and in vitro was inhibited by USP9X inhibitor. Cell migration and invasion were significantly inhibited by down-regulation of USP9X. USP9X was involved in extracellular signal-regulated kinase 1/2 (ERK1/2) and phosphatidylinositol-3-kinases/protein-serine-threonine kinase (PI3K/Akt) pathway in osteosarcoma cells. Proliferation, migration and invasion of osteosarcoma cells were inhibited by down-regulation of USP9X, and were related to the ERK1/2 and PI3K/Akt signaling pathways, therefore, it might probably become a new target for the prevention and treatment of osteosarcoma.
泛素特异性肽酶 9X（USP9X）已被报道与多种恶性肿瘤的发生和发展密切相关。然而，USP9X 如何参与骨肉瘤的发生和发展的机制尚未得到明确研究。本研究旨在探讨 USP9X 对骨肉瘤细胞增殖、迁移和侵袭的影响。
本研究招募了 67 名经组织学明确诊断为骨肉瘤的患者。采用骨肉瘤样本和细胞系来反映 USP9X 的表达水平。通过噻唑蓝（MTT）测定分析细胞增殖。通过 Transwell 实验和划痕愈合实验验证细胞迁移和侵袭能力。通过体内实验研究 USP9X 的作用。通过 Western blot 和定量实时 PCR（qRT-PCR）检测 USP9X 相关通路蛋白。
结果显示，USP9X 在骨肉瘤中的表达高于相邻组织。USP9X 阴性患者的总生存率优于 USP9X 阳性患者。USP9X 抑制剂抑制骨肉瘤细胞在体内和体外的生长。下调 USP9X 显著抑制细胞迁移和侵袭。USP9X 参与骨肉瘤细胞中细胞外信号调节激酶 1/2（ERK1/2）和磷脂酰肌醇-3-激酶/蛋白丝氨酸苏氨酸激酶（PI3K/Akt）通路。下调 USP9X 抑制骨肉瘤细胞的增殖、迁移和侵袭，与 ERK1/2 和 PI3K/Akt 信号通路有关，因此，USP9X 可能成为预防和治疗骨肉瘤的新靶点。
