Yang Fan, Wang Mao
Department of Bone and Soft-Tissue Tumor, Shanxi Province Cancer Hospital/ Shanxi Hospital Affiliated to Cancer Hospital, Chinese Academy of Medical Sciences/Cancer Hospital Affiliated to Shanxi Medical University, Taiyuan City, Shanxi Province 030013, China.
J Bone Oncol. 2025 Jul 12;53:100701. doi: 10.1016/j.jbo.2025.100701. eCollection 2025 Aug.
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.
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. assay was carried out using xenograft models in mice.
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. , NAT10/ITGB3 axis also promoted OS tumor growth in mice.
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.
N-乙酰转移酶10（NAT10）在肿瘤进展（包括骨肉瘤（OS））中作为N4-乙酰胞苷（ac4C）修饰的“书写者”。其在骨肉瘤中的分子机制仍不完全清楚。本研究旨在揭示骨肉瘤中与阴阳1蛋白（YY1）和整合素β3（ITGB3）相关的NAT10的上下游机制。
通过实时定量PCR和蛋白质印迹法检测基因mRNA和蛋白质水平。进行细胞计数试剂盒-8、EdU检测、流式细胞术/TUNEL染色检测、Transwell检测和划痕检测，以评估细胞活力、增殖、凋亡、侵袭和迁移。通过ac4C RNA免疫沉淀（ac4c RIP）、RIP、染色质免疫沉淀和双荧光素酶报告基因检测完成相互作用分析。使用小鼠异种移植模型进行检测。
骨肉瘤组织和细胞中NAT10表达较高。在NAT10沉默的骨肉瘤细胞中，细胞增殖、侵袭和迁移受到抑制，但凋亡增强。GSE237541数据集预测NAT10敲低后ITGB3受到抑制，PACES网站预测ITGB3中的ac4C位点。此外，发现NAT10可通过介导ac4C乙酰化上调ITGB3表达。ITGB3过表达挽救了由NAT10敲低引起的骨肉瘤细胞进展抑制。Jaspar预测YY1与NAT10启动子之间的结合。YY1可激活NAT10的转录调控以增加NAT10表达，YY1缺失通过降低NAT10表达阻断细胞恶性行为。更重要的是，YY1与NAT10相互作用以上调ITGB3表达。此外，NAT10/ITGB3轴也促进了小鼠骨肉瘤肿瘤的生长。
首次证实YY1调节NAT10的转录，首次表明NAT10介导ITGB3的ac4C修饰。YY1激活的NAT10可影响ITGB3，进而调节骨肉瘤的恶性发展。
