Department of Orthopedics, Shanghai University of Medicine and Health Sciences Affiliated Zhoupu Hospital, No. 1500, Zhouyuan Road, Pudong New District, Shanghai City, 201318, China.
Department of Orthopedics, Shanghai Public Health Clinical Center, No. 2901, Caolang Road, Jinshan District, Shanghai City, 201508, China.
J Orthop Surg Res. 2022 Feb 5;17(1):74. doi: 10.1186/s13018-022-02964-2.
Osteosarcoma is a type of bone malignancy that mainly occurred in teenagers. This investigation is aimed to clarify the effect of long non-coding RNA (lncRNA) LINC00662 on the proliferation, migration, and invasion in osteosarcoma and explore the underlying action mechanisms.
The mRNA expression of LINC00662 was determined by real-time quantitative polymerase chain reaction. Cell proliferation, migration, and invasion were evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, wound healing, and transwell assays, respectively. A dual-luciferase reporter assay was used to validate the target relationships Between microRNA (miR)-30b-3p and LINC00662/ ETS domain-containing protein 1 (ELK1). Western blotting was performed to determine the protein expression of ELK1. Xenograft model was established to evaluate the effects of LINC00662 silencing on tumor growth in vivo.
LncRNA LINC00662 and ELK1 were significantly increased, while miR-30b-3p was reduced in osteosarcoma tissues. The results of functional experiments indicated that transfection of small hairpin (sh)-LINC00662 and miR-30b-3p mimics repressed the migration, invasion, and proliferation of osteosarcoma cells. LncRNA LINC00662 also appeared to sponge miR-30b-3p in order to affect the expression of ELK1. Simultaneously, there were weak negative correlations between the expression of miR-30b-3p and LINC00662/ELK1 in osteosarcoma tissues. Rescue experiments suggested that ELK1 overexpression and downregulation of miR-30b-3p reversed the suppressive effects of sh-LINC00662 on the cell migration, invasion, and proliferation in osteosarcoma.
The current study indicated that knockdown of LINC00662 repressed cell migration, invasion, and proliferation through sponging miR-30b-3p to regulate the expression of ELK1 in osteosarcoma. These results may uncover a promising target for the treatment of osteosarcoma.
骨肉瘤是一种主要发生在青少年中的骨恶性肿瘤。本研究旨在阐明长链非编码 RNA（lncRNA）LINC00662 对骨肉瘤增殖、迁移和侵袭的影响，并探讨其潜在的作用机制。
采用实时定量聚合酶链反应测定 LINC00662 的 mRNA 表达。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐比色法、划痕愈合实验和 Transwell 实验分别评估细胞增殖、迁移和侵袭。双荧光素酶报告基因实验验证 microRNA（miR）-30b-3p 与 LINC00662/ETS 结构域蛋白 1（ELK1）之间的靶关系。Western blot 检测 ELK1 蛋白表达。建立异种移植模型评估体内沉默 LINC00662 对肿瘤生长的影响。
骨肉瘤组织中 LncRNA LINC00662 和 ELK1 显著升高，而 miR-30b-3p 降低。功能实验结果表明，转染小发夹（sh）-LINC00662 和 miR-30b-3p 模拟物可抑制骨肉瘤细胞的迁移、侵袭和增殖。LINC00662 似乎通过海绵吸附 miR-30b-3p 来影响 ELK1 的表达。同时，骨肉瘤组织中 miR-30b-3p 与 LINC00662/ELK1 的表达呈弱负相关。挽救实验表明，ELK1 过表达和 miR-30b-3p 下调逆转了 sh-LINC00662 对骨肉瘤细胞迁移、侵袭和增殖的抑制作用。
本研究表明，敲低 LINC00662 通过海绵吸附 miR-30b-3p 调节 ELK1 的表达，抑制骨肉瘤细胞的迁移、侵袭和增殖。这些结果可能为骨肉瘤的治疗提供一个有前途的靶点。
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