长链非编码 RNA PVT1 通过海绵吸附 miR-484 促进骨肉瘤细胞转移。

Long noncoding RNA PVT1 promotes metastasis via miR-484 sponging in osteosarcoma cells.

机构信息

Department of Spinal Surgery, The First Hospital of Jilin University, Changchun, P.R. China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Mar;24(5):2229-2238. doi: 10.26355/eurrev_202003_20488.

DOI:10.26355/eurrev_202003_20488

PMID:32196583

Abstract

OBJECTIVE

Long noncoding RNAs (lncRNAs) are widely involved in various malignancies including osteosarcoma. In the current study, we aimed to illustrate the role of lncRNA plasmacytoma variant translocation 1 (PVT1) in osteosarcoma.

PATIENTS AND METHODS

Expression of PVT1 and microRNA-486 (miR-486) in osteosarcoma tissue specimens and cell lines were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assays and in situ hybridizations (ISH) assay. Transwell migration/invasion assays were performed to determine the metastatic ability changes in osteosarcoma cells. Kaplan-Meier survival analysis was applied to analyze the overall survival (OS) of patients with osteosarcoma. Luciferase assays were used to evaluate the targeted binding effect between PVT1 and miR-486.

RESULTS

We illustrated that lncRNA plasmacytoma variant translocation 1 (PVT1) was upregulated in osteosarcoma, and it was correlated with poor prognosis of patients with osteosarcoma. Furthermore, we found that PVT1, via constructed loss of function and gain of function assays, promoted osteosarcoma cells migration and invasion. Meanwhile, we demonstrated that microRNA-486 (miR-486) was involved in PVT1-induced migration and invasion. We also uncovered that miR-486 was downregulated in osteosarcoma tissue specimens and cell lines. Functionally, we showed that upregulation of miR-486 reversed the facilitative effect of PVT1 on osteosarcoma cells migration and invasion, and vice versa. Mechanically, we illustrated that PVT1 interacted with miR-486 in a reciprocal suppressed manner. Moreover, we found that miR-486 could target to PVT1 via Luciferase assay. Lastly, we proved that PVT1 promoted osteosarcoma cells migration and invasion through miR-486 sponging.

CONCLUSIONS

We demonstrated that PVT1, functioning as an oncogene, promotes osteosarcoma cells metastasis via miR-486 sponging. PVT1/miR-486 axis might be a novel target in the molecular treatment of osteosarcoma.

摘要

目的

长链非编码 RNA（lncRNA）广泛参与包括骨肉瘤在内的各种恶性肿瘤。在本研究中，我们旨在阐明浆细胞瘤变异易位 1（PVT1）lncRNA 在骨肉瘤中的作用。

患者和方法

通过定量实时聚合酶链反应（qRT-PCR）检测骨肉瘤组织标本和细胞系中 PVT1 和 microRNA-486（miR-486）的表达，并通过原位杂交（ISH）检测。Transwell 迁移/侵袭实验用于确定骨肉瘤细胞转移能力的变化。Kaplan-Meier 生存分析用于分析骨肉瘤患者的总生存率（OS）。荧光素酶实验用于评估 PVT1 和 miR-486 之间的靶向结合效应。

结果

我们表明，浆细胞瘤变异易位 1（PVT1）lncRNA 在骨肉瘤中上调，并与骨肉瘤患者的不良预后相关。此外，我们发现 PVT1 通过构建的功能丧失和功能获得实验促进了骨肉瘤细胞的迁移和侵袭。同时，我们证明 microRNA-486（miR-486）参与了 PVT1 诱导的迁移和侵袭。我们还发现 miR-486 在骨肉瘤组织标本和细胞系中下调。功能上，我们表明 miR-486 的上调逆转了 PVT1 对骨肉瘤细胞迁移和侵袭的促进作用，反之亦然。机制上，我们表明 PVT1 以相互抑制的方式与 miR-486 相互作用。此外，我们通过荧光素酶实验证明了 PVT1 可以通过 miR-486 靶向。最后，我们证明 PVT1 通过 miR-486 海绵作用促进骨肉瘤细胞迁移和侵袭。