Department of Orthopedics, People's Hospital of Rizhao, Affiliated to Clinical Hospital of Jining Medical University, 126 Tai'an Road, Xinshi District, Rizhao, 276800, Shandong, China.
People's Hospital of Rizhao, 126 Taian Rd, Donggang District, Rizhao, Shandong, China.
Hum Cell. 2021 Jul;34(4):1255-1265. doi: 10.1007/s13577-021-00536-w. Epub 2021 Apr 23.
Osteosarcoma (OSA), the malignant bone tumor, predominantly affecting children and adolescents, threatens the life and life quality of the patients. An increasing number of studies have indicated the role of long non-coding RNA (lncRNA) dysregulation in cancer biology. Herein, the study was aimed to explore the role of FGD5 antisense RNA 1 (FGD5-AS1), a lncRNA, in OSA. Expression levels of FGD5-AS1, miR-506-3p and RAB3D mRNA were quantified utilizing qRT-PCR. The expression of RAB3D protein was examined employing Western blot. A series of functional experiments including CCK-8 assay, BrdU assay, wound healing assay, Transwell assay were performed for studying the effects of FGD5-AS1 on the malignancy of OSA cell lines 143B and HOS. The binding site between miR-506-3p and FGD5-AS1 was identified and validated by luciferase reporter assay and RNA immunoprecipitation assay. It was demonstrated that the expression of FGD5-AS1 was up-regulated in OSA tissues and cell lines, and its high expression is associated with higher Enneking stage and poorer histological differentiation. Gain-of-function and loss-of-function studies suggested that FGD5-AS1 facilitated OSA cells proliferation and migration. The promoting effects of FGD5-AS1 overexpression on OSA cell proliferation and migration could be counteracted by miR-506-3p. Moreover, FGD5-AS1 competitively adsorbed miR-506-3p to repress its expression so as to up-regulate the expression of RAB3D. These results indicate that FGD5-AS1 is capable of expediting OSA cell proliferation and migration via sponging miR-506-3p to up-regulate RAB3D.
骨肉瘤（OSA）是一种主要影响儿童和青少年的恶性骨肿瘤，威胁着患者的生命和生活质量。越来越多的研究表明长链非编码 RNA（lncRNA）失调在癌症生物学中的作用。在此，本研究旨在探讨 lncRNA FGD5 反义 RNA 1（FGD5-AS1）在 OSA 中的作用。利用 qRT-PCR 定量检测 FGD5-AS1、miR-506-3p 和 RAB3D mRNA 的表达水平。采用 Western blot 检测 RAB3D 蛋白的表达。通过 CCK-8 assay、BrdU assay、划痕愈合 assay、Transwell assay 等一系列功能实验研究 FGD5-AS1 对 OSA 细胞系 143B 和 HOS 恶性表型的影响。通过荧光素酶报告基因 assay 和 RNA 免疫沉淀 assay 鉴定和验证 miR-506-3p 与 FGD5-AS1 之间的结合位点。结果表明，FGD5-AS1 在 OSA 组织和细胞系中表达上调，其高表达与更高的 Enneking 分期和较差的组织学分化相关。功能获得和功能丧失研究表明，FGD5-AS1 促进 OSA 细胞增殖和迁移。FGD5-AS1 过表达对 OSA 细胞增殖和迁移的促进作用可以被 miR-506-3p 逆转。此外，FGD5-AS1 通过竞争性吸附 miR-506-3p 来抑制其表达，从而上调 RAB3D 的表达。这些结果表明，FGD5-AS1 通过海绵吸附 miR-506-3p 来上调 RAB3D，从而加速 OSA 细胞的增殖和迁移。
