Department of Orthopedics and Traumatology, Zibo Hospital of Integrated Traditional Chinese and Western Medicine, Zibo, 255000, Shandong Province, China.
CT Imaging Department of Zibo Central Hospital, Zibo, 255000, Shandong Province, China.
World J Surg Oncol. 2020 May 1;18(1):83. doi: 10.1186/s12957-020-01859-y.
Osteosarcoma (OS) is a primary malignant bone tumor with a high rate of metastasis and a short 5-year survival rate. MiR-363 was downregulated in a variety of tumors and played a role in suppressing tumors. However, the roles of miR-363 in osteosarcoma remain unknown; thus, the purpose of this study was to explore the functions of miR-363 in osteosarcoma.
CCK-8 and transwell assays were performed to evaluate the proliferation, migration, and invasion abilities of MG63 cells. The epithelial-mesenchymal transition (EMT) and apoptosis-associated proteins were measured by using Western blot assay. Luciferase reporter assay was utilized to verify whether miR-363 directly bound to the 3'-UTR of NOB1 mRNA.
MiR-363 was downregulated while NOB1 was upregulated in osteosarcoma clinical tissue specimens and cell lines as compared with the adjacent normal tissue specimens and normal cell line. The miR-363 is reversely correlated with the expression of NOB1 in osteosarcoma tissues. Overexpression of miR-363 suppressed the ability of cell migration, invasion, and EMT, whereas low expression of miR-363 promoted this ability. In addition, miR-363 inhibited osteosarcoma proliferation both in vitro and in vivo and inhibited the apoptosis in MG63 cells. Interference of NOB1 could inhibit the migration, invasion, and EMT of osteosarcoma cell line MG63. NOB1 was verified to be a direct target of miR-363 and its expression was mediated by miR-363. Re-expression of NOB1 could partially reverse the inhibitory effect of miR-363 on cell migration and invasion. In addition, low expression of miR-363 or overexpression of NOB1 predicted poor prognosis of osteosarcoma patients.
MiR-363 inhibited osteosarcoma the proliferation, migration, invasion, and EMT and induced the apoptosis by directly targeting NOB1 in MG63 cells. The newly identified miR-363/NOB1 axis provides novel insights into the pathogenesis of osteosarcoma.
骨肉瘤（OS）是一种原发性恶性骨肿瘤，转移率高，5 年生存率短。miR-363 在多种肿瘤中下调，发挥抑制肿瘤的作用。然而，miR-363 在骨肉瘤中的作用尚不清楚；因此，本研究旨在探讨 miR-363 在骨肉瘤中的功能。
采用 CCK-8 和 Transwell 实验评估 MG63 细胞的增殖、迁移和侵袭能力。Western blot 实验检测上皮-间充质转化（EMT）和凋亡相关蛋白。荧光素酶报告实验验证 miR-363 是否直接结合到 NOB1 mRNA 的 3'-UTR。
与相邻正常组织标本和正常细胞系相比，骨肉瘤临床组织标本和细胞系中 miR-363 下调，而 NOB1 上调。miR-363 在骨肉瘤组织中与 NOB1 的表达呈负相关。过表达 miR-363 抑制细胞迁移、侵袭和 EMT 能力，而低表达 miR-363 促进这种能力。此外，miR-363 抑制骨肉瘤体外和体内的增殖并抑制 MG63 细胞的凋亡。NOB1 的干扰可抑制骨肉瘤细胞系 MG63 的迁移、侵袭和 EMT。NOB1 被验证为 miR-363 的直接靶标，其表达受 miR-363 介导。NOB1 的重新表达可部分逆转 miR-363 对细胞迁移和侵袭的抑制作用。此外，miR-363 低表达或 NOB1 过表达预示着骨肉瘤患者的预后不良。
miR-363 通过直接靶向 MG63 细胞中的 NOB1 抑制骨肉瘤的增殖、迁移、侵袭和 EMT，并诱导凋亡。新鉴定的 miR-363/NOB1 轴为骨肉瘤的发病机制提供了新的见解。
