Knockdown of lncRNA HOXA-AS2 Inhibits Viability, Migration and Invasion of Osteosarcoma Cells by miR-124-3p/E2F3
Knockdown of lncRNA HOXA-AS2 Inhibits Viability, Migration and Invasion of Osteosarcoma Cells by miR-124-3p/E2F3
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DOI：
10.2147/ott.s220072
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发表时间：
2019-01-01
影响因子：
4
通讯作者：
Zhang, Xinhua
中科院分区：
文献类型：
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作者：
Wang, Linyi;Wang, Lijuan;Zhang, Xinhua
背景：骨肉瘤（OS）是最常见的骨恶性肿瘤之一。长非编码 RNA (lncRNA) 已被证明参与许多癌症，包括 OS。本研究旨在探讨lncRNA同源盒A簇反义RNA2（HOXA-A52）的生物学功能及其在OS进展中的潜在机制。方法：本研究招募了27名OS患者。培养 U2OS 和 MG-63 细胞用于体外分析。通过实时定量聚合酶链式反应或蛋白质印迹测量 HOXA-A52、microRNA-124-3p (miR-124-3p) 和 E2F 转录因子 3 (E2F3) 的水平。使用细胞计数试剂盒 8 或跨孔测定通过细胞活力、迁移和侵袭来研究 OS 进展。通过生物信息学分析、荧光素酶报告基因测定、RNA 免疫沉淀和生物素化 RNA Pull-down 探索 HOXA-A52、miR-124-3p 和 E2F3 之间的相互作用。通过将U2OS细胞注射到裸鼠体内建立异种移植模型。结果：OS组织和细胞中HOXA-AS2表达增加，并与患者的不良生存相关。 HOXA-AS2 的敲低抑制了 OS 细胞的活力、迁移和侵袭。 miR-124-3p 可以与 HOXA-AS2 结合，其缺陷逆转了 HOXA-AS2 敲低的抑制作用。此外，E2F3 作为 miR-124-3p 的靶标并受到 HOXA-A52 的正向调节。 E2F3 的沉默抑制了 OS 进展，而这种进展被 miR-124-3p 耗竭所消除。 HOXA-AS2 的干扰通过上调 miR-124-3p 和下调 E2F3 来减弱 U2OS 异种移植肿瘤的生长。结论：HOXA-AS2 沉默可能通过作为 miR-124-3p 靶向 E2F3 的诱饵来阻碍 OS 进展，这表明 HOXA-AS2 作为 OS 的有希望的治疗靶点的新证据。
Background: Osteosarcoma (OS) is one of the most frequent bone malignancies. Long noncoding RNAs (lncRNAs) have been revealed to participate in many cancers, including OS. This study aimed to explore the biological function of lncRNA homeobox A cluster antisense RNA2 (HOXA-A52) and its potential mechanism in OS progression.Methods: Twenty-seven OS patients were recruited for this study. U2OS and MG-63 cells were cultured for in vitro analyses. The levels of HOXA-A52, microRNA-124-3p (miR-124-3p) and E2F transcription factor 3 (E2F3) were measured by quantitative real-time polymerase chain reaction or Western blot. OS progression was investigated by cell viability, migration and invasion using cell counting kit-8 or trans-well assay. The interaction among HOXA-A52, miR-124-3p and E2F3 was explored by bioinformatics analysis, luciferase reporter assay, RNA immunoprecipitation and biotinylated RNA pull-down. Xenograft model was established by injecting U2OS cells into nude mice.Results: HOXA-AS2 expression was increased in OS tissues and cells and associated with poor survival of patients. Knockdown of HOXA-AS2 inhibited cell viability, migration and invasion in OS cells. miR-124-3p could bind with HOXA-AS2 and its deficiency reversed the suppressive role of HOXA-AS2 knockdown. Moreover, E2F3 acted as a target of miR-124-3p and positively regulated by HOXA-A52. Silence of E2F3 suppressed OS progression, which was abolished by miR-124-3p exhaustion. Interference of HOXA-AS2 attenuated U2OS xenograft tumor growth via upregulating miR-124-3p and downregulating E2F3.Conclusion: HOXA-AS2 silence impeded OS progression possibly by functioning as a decoy of miR-124-3p to target E2F3, indicating novel evidence of HOXA-AS2 as a promising therapeutic target of OS.