阴阳1蛋白激活的n-乙酰转移酶10通过整合素β3的ac4C乙酰化驱动骨肉瘤细胞恶性进展

IF 3.5 2区医学 Q2 Medicine

Journal of Bone Oncology Pub Date : 2025-07-12 DOI:10.1016/j.jbo.2025.100701

Fan Yang, Mao Wang

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引用次数: 0

摘要

n -乙酰基转移酶10 （NAT10）在包括骨肉瘤（OS）在内的肿瘤进展中起着n4 -乙酰胞苷（ac4C）修饰的“作者”作用。其在OS中的分子机制尚不完全清楚。本研究旨在揭示在OS中与阴阳1蛋白（YY1）和整合素β3 （ITGB3）相关的NAT10的上下游机制。方法采用实时定量PCR和Western blotting检测各组小鼠基因mRNA和蛋白表达水平。采用细胞计数试剂盒-8、EdU法、流式细胞术/TUNEL染色法、transwell法和划痕法评估细胞活力、增殖、凋亡、侵袭和迁移。通过ac4C RNA免疫沉淀（ac4C RIP）、RIP、染色质IP和双荧光素酶报告基因测定完成相互作用分析。采用小鼠异种移植物模型进行体内实验。结果6个组织和细胞均高表达NAT10。在nat10沉默的OS细胞中，细胞增殖、侵袭和迁移受到抑制，但细胞凋亡增强。GSE237541数据集预测了NAT10敲低后ITGB3的抑制作用，PACES网站预测了ITGB3中的ac4C位点。进一步发现NAT10可以通过介导ac4C乙酰化上调ITGB3的表达。ITGB3过表达可避免NAT10敲低引起的OS细胞进展抑制。Jaspar预测了YY1与NAT10启动子之间的结合。YY1可以激活NAT10的转录调控，增加NAT10的表达，YY1缺失通过降低NAT10的表达来阻断细胞的恶性行为。更重要的是，YY1与NAT10相互作用，上调ITGB3的表达。在体内，NAT10/ITGB3轴也促进了小鼠OS肿瘤的生长。结论yy1首次被证实调控NAT10的转录，NAT10首次被证实介导ITGB3的ac4C修饰。yy1激活的NAT10可影响ITGB3，进而调控OS的恶性发展。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Yin Yang 1 protein-activated N-acetyltransferase 10 drives cell malignant progression of osteosarcoma through ac4C acetylation of integrin β3

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Yin Yang 1 protein-activated N-acetyltransferase 10 drives cell malignant progression of osteosarcoma through ac4C acetylation of integrin β3

Background

N-acetyltransferase 10 (NAT10) acts as the “writer” of N4-acetylcytidine (ac4C) modification in tumor progression, including osteosarcoma (OS). Its molecular mechanism in OS remains not fully clear. This study endeavored to disclose the upstream and downstream mechanism of NAT10 related to Yin Yang 1 protein (YY1) and integrin β3 (ITGB3) in OS.

Methods

Gene mRNA and protein levels were assayed via real-time quantitative PCR and Western blotting. Cell counting kit-8, EdU assay, flow cytometry/TUNEL staining assay, transwell assay, and scratch assay were conducted to assess cell viability, proliferation, apoptosis, invasion, and migration. Interaction analysis was completed through ac4C RNA immunoprecipitation (ac4c RIP), RIP, chromatin IP and dual-luciferase reporter assay. In vivo assay was carried out using xenograft models in mice.

Results

OS tissues and cells showed the high expression of NAT10. Cell proliferation, invasion, and migration were suppressed but apoptosis was enhanced in NAT10-silenced OS cells. GSE237541 dataset has predicted the inhibition of ITGB3 after NAT10 knockdown, and PACES website predicted ac4C site in ITGB3. Furthermore, it was found that NAT10 could up-regulate ITGB3 expression by mediating ac4C acetylation. ITGB3 overexpression recused OS cell progression inhibition caused by NAT10 knockdown. Jaspar predicted the binding between YY1 and NAT10 promoter. YY1 could activate the transcriptional regulation of NAT10 to increase NAT10 expression, and YY1 depletion blocked cell malignant behaviors via reducing NAT10 expression. More importantly, YY1 interacted with NAT10 to up-regulate ITGB3 expression. In vivo, NAT10/ITGB3 axis also promoted OS tumor growth in mice.

Conclusion

YY1 was firstly affirmed to regulate transcription of NAT10, and NAT10 was firstly indicated to mediate ac4C modification of ITGB3. YY1-activated NAT10 could affect ITGB3 and then modulated the malignant development of OS.

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来源期刊

Journal of Bone Oncology ONCOLOGY-

CiteScore

7.20

自引率

2.90%

发文量

审稿时长

34 days

期刊介绍： The Journal of Bone Oncology is a peer-reviewed international journal aimed at presenting basic, translational and clinical high-quality research related to bone and cancer. As the first journal dedicated to cancer induced bone diseases, JBO welcomes original research articles, review articles, editorials and opinion pieces. Case reports will only be considered in exceptional circumstances and only when accompanied by a comprehensive review of the subject. The areas covered by the journal include: Bone metastases (pathophysiology, epidemiology, diagnostics, clinical features, prevention, treatment) Preclinical models of metastasis Bone microenvironment in cancer (stem cell, bone cell and cancer interactions) Bone targeted therapy (pharmacology, therapeutic targets, drug development, clinical trials, side-effects, outcome research, health economics) Cancer treatment induced bone loss (epidemiology, pathophysiology, prevention and management) Bone imaging (clinical and animal, skeletal interventional radiology) Bone biomarkers (clinical and translational applications) Radiotherapy and radio-isotopes Skeletal complications Bone pain (mechanisms and management) Orthopaedic cancer surgery Primary bone tumours Clinical guidelines Multidisciplinary care Keywords: bisphosphonate, bone, breast cancer, cancer, CTIBL, denosumab, metastasis, myeloma, osteoblast, osteoclast, osteooncology, osteo-oncology, prostate cancer, skeleton, tumour.