ac4C-seq in NAT10-knockdown PANC-1 cells

Pancreatic cancer is a lethal diease with high tendency of metastasis. Howerver, the mechanisms of pancreatic cancer are sitill unclear. To explore the roles of N4-acetylation (ac4C) RNA modification and its involved N-Acetyltransferase 10 (NAT10) in pancreatic ductal adenocarcinoma (PDAC), we performed profiling by high throughput sequencing. In this study, we investigate the effects of NAT10 knockdown on N4-acetylcytidine (ac4C) modification in mRNA within PANC-1 cells using ac4C-seq. By employing RNA interference to specifically knock down NAT10 expression in PANC-1 cells, we aim to elucidate its impact on ac4C RNA modifications, which have been implicated in various cellular processes and cancer progression. Total RNA was extracted and mRNA was captured and treated with sodium borohydride (NaBH4) for detection of ac4C sites.Following library preparation, sequencing was performed on an Illumina Novaseq 6000 platform. Bioinformatics analyses identified significant changes in ac4C modification patterns due to NAT10 depletion. This dataset provides a valuable resource for further exploration of ac4C modifications in mRNA and their role in PDAC. NAT10 knockdown or ctrl PANC-1 cells were used to investigate the genes with changes in ac4C sites regulated by NAT10.

胰腺癌是一种致死性极强且极易发生转移的恶性肿瘤，但其发病机制仍未明确。为探究N4-乙酰化（N4-acetylation）RNA修饰及其相关的N-乙酰基转移酶10（NAT10）在胰腺导管腺癌（PDAC）中的作用，我们通过高通量测序开展了谱型分析。本研究利用ac4C-seq技术，在PANC-1细胞中探究了NAT10敲低对信使RNA（mRNA）上N4-乙酰胞苷（ac4C）修饰的影响。我们通过RNA干扰技术特异性敲低PANC-1细胞中NAT10的表达，旨在阐明其对ac4C RNA修饰的调控作用——此类修饰已被证实参与多种细胞进程及肿瘤进展过程。我们提取总RNA，对mRNA进行富集捕获后，采用硼氢化钠（NaBH4）处理以检测ac4C修饰位点。完成文库构建后，我们在Illumina Novaseq 6000测序平台上完成了测序。生物信息学分析结果显示，NAT10表达敲低会显著改变ac4C的修饰模式。本数据集为进一步探究mRNA上的ac4C修饰及其在PDAC中的作用提供了宝贵的研究资源。本研究采用NAT10敲低组与对照组PANC-1细胞，旨在筛选受NAT10调控的ac4C位点发生改变的基因。