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METTL3 介导的 circRNF220 的 m6A 修饰调节 miR-330-5p/survivin 轴促进骨肉瘤进展。

METTL3-mediated m6A modification of circRNF220 modulates miR-330-5p/survivin axis to promote osteosarcoma progression.

机构信息

Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan, 430060, China.

出版信息

J Cancer Res Clin Oncol. 2023 Dec;149(19):17347-17360. doi: 10.1007/s00432-023-05455-x. Epub 2023 Oct 14.

Abstract

BACKGROUND

Circular RNAs (circRNAs) play a crucial role in regulating various physiological processes. However, the precise regulatory mechanisms of circRNF220s in osteosarcoma (OS) are not well understood.

METHODS

The abundances of circRNF220, miR-330-5p, and survivin were determined using qRT-PCR. To assess the m6A accumulation in circRNF220, a methylated RNA immunoprecipitation (Me-RIP) assay was conducted. Cellular multiplication, motility, and invasion were examined using the cell Counting Kit-8 (CCK-8), EdU, colony formation, Transwell, and wound-healing assays. The binding relationships were measured through RNA immunoprecipitation (RIP) and luciferase reporter assays. In vivo functionality was assessed using xenograft models.

RESULTS

CircRNF220 was identified as being overexpressed in both OS cells and tissues. In vitro experiments demonstrated that silencing circRNF220 impeded the proliferation, invasion, and motility of OS cells. Similarly, in vivo studies confirmed that downregulating circRNF220 inhibited the growth of OS. Further mechanistic investigations unveiled that METTL3-modulated circRNF220 regulated the progression of OS by upregulating survivin expression through acting as a sponge for miR-330-5p.

CONCLUSION

The modulation of METTL3-regulated circRNF220 has been found to promote the progression of OS by modulating the miR-330-5p/survivin axis. This novel finding suggests a potentially unique approach to managing OS.

摘要

背景

环状 RNA(circRNAs)在调控多种生理过程中发挥着关键作用。然而,circRNF220 在骨肉瘤(OS)中的精确调控机制尚不清楚。

方法

使用 qRT-PCR 测定 circRNF220、miR-330-5p 和 survivin 的丰度。为了评估 circRNF220 中的 m6A 积累,进行了甲基化 RNA 免疫沉淀(Me-RIP)测定。使用细胞计数试剂盒-8(CCK-8)、EdU、集落形成、Transwell 和划痕愈合测定来评估细胞增殖、迁移和侵袭。通过 RNA 免疫沉淀(RIP)和荧光素酶报告基因测定来测量结合关系。使用异种移植模型评估体内功能。

结果

circRNF220 在 OS 细胞和组织中均被鉴定为过度表达。体外实验表明,沉默 circRNF220 抑制了 OS 细胞的增殖、侵袭和迁移。同样,体内研究证实下调 circRNF220 抑制了 OS 的生长。进一步的机制研究揭示,METTL3 调节的 circRNF220 通过作为 miR-330-5p 的海绵来上调 survivin 表达,从而调节 OS 的进展。

结论

METTL3 调节的 circRNF220 的调节被发现通过调节 miR-330-5p/survivin 轴促进 OS 的进展。这一新发现为 OS 的管理提供了一种潜在的独特方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc62/11797187/4f59af719e88/432_2023_5455_Fig1_HTML.jpg

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