Abstract：
BACKGROUND: N-acetyltransferase 10 (NAT10), the only RNA cytosine acetyltransferase known in humans, contributes to cancer tumorigenesis and progression. This study aims to investigate the effect of NAT10 on the malignant biological properties of gastric cancer (GC) and its underlying mechanism.
METHODS: The expression and prognostic significance of NAT10 in GC were analyzed using The Cancer Genome Atlas (TCGA) and Sun Yat-sen University (SYSU) cohorts. The influence of NAT10 on the malignant biological behaviors of GC was detected by Cell Counting Kit-8 (CCK-8) assay, plate colony formation assay, 5-ethynyl-2\'-deoxyuridine (EdU), Transwell migration and invasion assays, scratch wound assay, flow cytometric analysis, and animal studies. The overall level of N4 acetylcytidine (ac4C) in GC was detected by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The downstream signal pathways of NAT10 were analyzed by Gene Set Enrichment Analysis (GSEA) and verified by Western blot (WB) and immunofluorescence (IF).
RESULTS: The significant upregulation of NAT10 expression in GC was associated with a poor prognosis. The knockdown of NAT10 markedly suppressed GC cell proliferation, migration, invasion, and cell cycle progression. Downregulating NAT10 reduced ac4C levels and inhibited AKT phosphorylation and epithelial-mesenchymal transition (EMT) in GC.
CONCLUSIONS: NAT10 functions as an oncogene and may provide a new therapeutic target in GC.
METHODS: The expression and prognostic significance of NAT10 in GC were analyzed using The Cancer Genome Atlas (TCGA) and Sun Yat-sen University (SYSU) cohorts. The influence of NAT10 on the malignant biological behaviors of GC was detected by Cell Counting Kit-8 (CCK-8) assay, plate colony formation assay, 5-ethynyl-2\'-deoxyuridine (EdU), Transwell migration and invasion assays, scratch wound assay, flow cytometric analysis, and animal studies. The overall level of N4 acetylcytidine (ac4C) in GC was detected by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The downstream signal pathways of NAT10 were analyzed by Gene Set Enrichment Analysis (GSEA) and verified by Western blot (WB) and immunofluorescence (IF).
RESULTS: The significant upregulation of NAT10 expression in GC was associated with a poor prognosis. The knockdown of NAT10 markedly suppressed GC cell proliferation, migration, invasion, and cell cycle progression. Downregulating NAT10 reduced ac4C levels and inhibited AKT phosphorylation and epithelial-mesenchymal transition (EMT) in GC.
CONCLUSIONS: NAT10 functions as an oncogene and may provide a new therapeutic target in GC.
摘要：
背景：N-乙酰转移酶10（NAT10），人类唯一已知的RNA胞嘧啶乙酰转移酶，有助于癌症的发生和发展。本研究旨在探讨NAT10对胃癌恶性生物学特性的影响及其机制。
方法：使用癌症基因组图谱（TCGA）和中山大学（SYSU）队列分析NAT10在GC中的表达和预后意义。用细胞计数试剂盒(CCK-8)法检测NAT10对GC恶性生物学行为的影响,平板集落形成试验，5-乙炔基-2'-脱氧尿苷（EdU），Transwell迁移和入侵测定，划痕伤口试验，流式细胞仪分析,和动物研究。通过液相色谱-串联质谱(LC-MS/MS)检测GC中N4乙酰胞苷(ac4C)的总体水平。NAT10的下游信号通路通过基因集富集分析（GSEA）进行分析，并通过蛋白质印迹（WB）和免疫荧光（IF）进行验证。
结果：GC中NAT10表达的显著上调与不良预后相关。NAT10的敲减显著抑制GC细胞增殖，迁移,入侵,和细胞周期进程。下调NAT10可降低ac4C水平并抑制GC中AKT磷酸化和上皮-间质转化（EMT）。
结论：NAT10作为癌基因发挥作用，可能为GC提供新的治疗靶点。
方法：使用癌症基因组图谱（TCGA）和中山大学（SYSU）队列分析NAT10在GC中的表达和预后意义。用细胞计数试剂盒(CCK-8)法检测NAT10对GC恶性生物学行为的影响,平板集落形成试验，5-乙炔基-2'-脱氧尿苷（EdU），Transwell迁移和入侵测定，划痕伤口试验，流式细胞仪分析,和动物研究。通过液相色谱-串联质谱(LC-MS/MS)检测GC中N4乙酰胞苷(ac4C)的总体水平。NAT10的下游信号通路通过基因集富集分析（GSEA）进行分析，并通过蛋白质印迹（WB）和免疫荧光（IF）进行验证。
结果：GC中NAT10表达的显著上调与不良预后相关。NAT10的敲减显著抑制GC细胞增殖，迁移,入侵,和细胞周期进程。下调NAT10可降低ac4C水平并抑制GC中AKT磷酸化和上皮-间质转化（EMT）。
结论：NAT10作为癌基因发挥作用，可能为GC提供新的治疗靶点。
