Wu Hanhua, Xu Hua, Man Yunan, Huang Piwei, Huang Linhai, He Maolin
Division of Spinal Surgery, The First Affiliated Hospital of Guangxi Medical University, Shuangyong Road 6, Nanning, Guangxi Zhuang Autonomous Region, 530021, PR China.
Center for Education Evaluation & Faculty Development, Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530021, PR China.
Biochem Biophys Res Commun. 2025 Mar 25;754:151491. doi: 10.1016/j.bbrc.2025.151491. Epub 2025 Feb 14.
Osteosarcoma (OS) is the most common primary malignant bone neoplasm in children and adolescents, characterized by high mortality and disability owing to frequent relapse and metastasis. However, N-alpha-acetyltransferase 40 (NAA40) molecular mechanisms underlying OS progression and metastasis remain unexplored.
Bioinformatics analysis was used to evaluate NAA40 role in OS data from GEO and TARGET database. OS cell multiplication, invasion and migration were gauged in CCK8, EdU assays, and Transwell assays. RT-qPCR, ChIP-qPCR, dual luciferase reporter assay and rescue experiments were to explore NAA40 regulatory mechanism. Animal experiments further confirmed cell-based assays and NAA40 molecular mechanism.
Herein NAA40 expression was upregulated in OS samples and associated with shorter survival among patients. Functionally, NAA40 depletion resulted in reduced OS cell viability, decreased migration, and invasion in vitro. Mechanistically, NAA40 loss was associated with increased H4S1ph and H4R3me2a and decreased H4R3me2s.NAA40 overexpression improved the transcriptional activity in the promoter of AGR2. Histone marks, H3K4me3 and H3K27me3, at the AGR2 promoter were altered, inducing changes in AGR2 expression in NAA40-depleted OS cells. Anterior gradient 2 (AGR2) was identified as a downstream target of NAA40.AGR2 knockdown in OS cells resulted in reduced viability, decreased migration, and invasion. Ectopic overexpression of AGR2 partially rescued these phenotypic changes. In vivo experiments revealed that NAA40 depletion led to reduced AGR2 protein levels, inhibiting the proliferative and metastatic potential of OS cells.
NAA40 contributes to OS development and progression by epigenetically regulating AGR2 expression.
骨肉瘤（OS）是儿童和青少年中最常见的原发性恶性骨肿瘤，因其频繁复发和转移导致高死亡率和致残率。然而，N-α-乙酰转移酶40（NAA40）在骨肉瘤进展和转移中的分子机制仍未被探索。
利用生物信息学分析评估NAA40在来自GEO和TARGET数据库的骨肉瘤数据中的作用。通过CCK8、EdU实验和Transwell实验检测骨肉瘤细胞的增殖、侵袭和迁移。采用RT-qPCR、ChIP-qPCR、双荧光素酶报告基因实验和拯救实验来探索NAA40的调控机制。动物实验进一步证实了基于细胞的实验结果和NAA40的分子机制。
在此研究中，NAA40在骨肉瘤样本中表达上调，且与患者较短的生存期相关。在功能上，NAA40缺失导致骨肉瘤细胞活力降低，体外迁移和侵袭能力下降。在机制上，NAA40缺失与H4S1ph和H4R3me2a增加以及H4R3me2s减少有关。NAA40过表达提高了AGR2启动子的转录活性。AGR2启动子处的组蛋白标记H3K4me3和H3K27me3发生改变，导致NAA40缺失的骨肉瘤细胞中AGR2表达发生变化。前梯度蛋白2（AGR2）被确定为NAA40的下游靶点。在骨肉瘤细胞中敲低AGR2导致细胞活力降低、迁移和侵袭能力下降。AGR2的异位过表达部分挽救了这些表型变化。体内实验表明，NAA40缺失导致AGR2蛋白水平降低，抑制了骨肉瘤细胞的增殖和转移潜能。
NAA40通过表观遗传调控AGR2的表达促进骨肉瘤的发生和发展。
