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环状 RNA PVT1 通过介导 miR-26b-5p/CCNB1 轴促进骨肉瘤的发生和转移。

CircPVT1 promotes the tumorigenesis and metastasis of osteosarcoma via mediation of miR-26b-5p/CCNB1 axis.

机构信息

Department of Pediatric Orthopedics, Hunan Children's Hospital, No. 86, Ziyuan Road, Changsha, 410007, Hunan, People's Republic of China.

出版信息

J Bone Miner Metab. 2022 Jul;40(4):581-593. doi: 10.1007/s00774-022-01326-6. Epub 2022 Jun 1.

Abstract

INTRODUCTION

Osteosarcoma (OS) is the most aggressive malignancy among the bone tumors in the world. Circular RNAs (circRNAs) have been reported to be participated in multiple cancers, including OS. Meanwhile, circPVT1 has been proved to be upregulated in OS. However, the mechanism by which circPVT1 mediates the tumorigenesis of OS remains to be further explored.

MATERIALS AND METHODS

Protein and gene expressions in OS cells were measured by western blot and RT-qPCR, respectively. Cell growth was assessed by flow cytometry and colony formation, respectively. In addition, cell migration was assessed by wound healing, and invasion was evaluated by Transwell assay. Meanwhile, the correlation among circPVT1, miR-26b-5p and CCNB1 was explored by RNA pull-down and dual luciferase assay. Finally, in vivo model was established to explore the role of circPVT1 in OS in vivo.

RESULTS

CircPVT1 and CCNB1 were significantly upregulated in OS cells, while miR-26b-5p was downregulated. Knockdown of circPVT1 notably inhibited proliferation and induced apoptosis of OS cells. CircPVT1 shRNA significantly suppressed the OS cell invasion and migration. Meanwhile, circPVT1 sponged miR-26b-5p and CCNB1 was found to be the direct target of miR-26b-5p. Furthermore, silencing of circPVT1 inhibited the growth and metastasis of OS in vivo.

CONCLUSION

Silencing of circPVT1 notably suppressed the tumorigenesis and metastasis of OS via miR-26b-5p/CCNB1 axis. Therefore, circPVT1 might be used as a target for OS treatment.

摘要

简介

骨肉瘤(OS)是世界范围内最具侵袭性的骨肿瘤恶性肿瘤。环状 RNA(circRNA)已被报道参与多种癌症,包括骨肉瘤。同时,circPVT1 已被证明在骨肉瘤中上调。然而,circPVT1 介导骨肉瘤发生的机制仍有待进一步探索。

材料与方法

通过 Western blot 和 RT-qPCR 分别测量 OS 细胞中的蛋白和基因表达。通过流式细胞术和集落形成分别评估细胞生长。此外,通过划痕愈合评估细胞迁移,通过 Transwell 测定评估细胞侵袭。同时,通过 RNA 下拉和双荧光素酶报告基因实验探索 circPVT1、miR-26b-5p 和 CCNB1 之间的相关性。最后,建立体内模型以研究 circPVT1 在体内骨肉瘤中的作用。

结果

circPVT1 和 CCNB1 在骨肉瘤细胞中显著上调,而 miR-26b-5p 下调。circPVT1 的敲低显著抑制 OS 细胞的增殖并诱导其凋亡。circPVT1 shRNA 显著抑制 OS 细胞的侵袭和迁移。同时,circPVT1 海绵吸附 miR-26b-5p,CCNB1 被发现是 miR-26b-5p 的直接靶标。此外,沉默 circPVT1 抑制了骨肉瘤在体内的生长和转移。

结论

沉默 circPVT1 通过 miR-26b-5p/CCNB1 轴显著抑制骨肉瘤的发生和转移。因此,circPVT1 可能作为骨肉瘤治疗的靶点。

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