N-乙酰基转移酶10通过ZEB1/MT1-MMP轴促进胰腺癌进展。

N-acetyltransferase 10 Promotes Pancreatic Cancer Progression Through ZEB1/MT1-MMP Axis.

作者信息

Wang Fangxia, Hu Yumeng, Zhang Shaobo, Ye Yuxin, Qi Haoran, Xu Yan, Zhang Hui, Liu Mingyang

机构信息

State Key Laboratory of Molecular Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital.

Department of Pathology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing.

出版信息

Pancreas. 2025 Sep 1;54(8):e674-e683. doi: 10.1097/MPA.0000000000002492.

DOI:10.1097/MPA.0000000000002492

PMID:40828650

Abstract

OBJECTIVES

To elucidate the role of N-acetyltransferase 10 (NAT10) in pancreatic cancer (PC) progression and its epigenetic mechanisms, particularly in relation to metastasis.

METHODS

TCGA and GTEx databases were used to analyze the expression and roles of NAT10 in pancreatic cancer. We constructed stable cell lines with NAT10 knockdown in PC cell lines, AsPC-1 and KPC. CCK-8, EdU assay, and colony formation assay were conducted to evaluate the capability of cell proliferation and clonogenesis in vitro. Meanwhile, a transwell assay was performed to assess the impact on invasion and metastasis abilities. The correlation between NAT10 and ZEB1 expression was verified by correlation analysis. The underlying mechanisms through which NAT10 regulates ZEB1 were confirmed by qPCR, western blot, RIP-qPCR, dot plot, and mRNA stability assay. Furthermore, the interplay among NAT10, ZEB1, and MT1-MMP was confirmed using similar experimental approaches. Rescue experiments involving ZEB1 overexpression further verified the role of NAT10/ZEB1/MT1-MMP axis in PC metastasis. In addition, the NAT10 inhibitor Remodelin was employed in a nude orthotopic PC model to investigate its effects on metastasis in vivo.

RESULTS

NAT10 was found to be upregulated in PC and was significantly associated with poor prognosis. After NAT10 knockdown, the ability of proliferation and metastasis of AsPC-1 and KPC was remarkably impaired, the degree of ac4C modification was decreased, and the mRNA stability of ZEB1 declined. Correlation analysis indicated a positive correlation among NAT10, ZEB1, and MT1-MMP, and the results of qPCR and western blot also verified this conclusion. Moreover, ZEB1 overexpression could significantly reverse the inhibition of migration and invasion induced by NAT10 depletion in AsPC-1. NAT10 inhibitor Remodelin treatment could reduce the degree of peritoneal and liver metastases in vivo.

CONCLUSIONS

Our study highlights the pivotal functions of NAT10 in the progression of PC and reveals the underlying epigenetic mechanism that NAT10 promotes metastasis via ZEB1/MT1-MMP axis.

摘要

目的

阐明N - 乙酰转移酶10（NAT10）在胰腺癌（PC）进展中的作用及其表观遗传机制，特别是与转移相关的机制。

方法

利用TCGA和GTEx数据库分析NAT10在胰腺癌中的表达及作用。我们在PC细胞系AsPC - 1和KPC中构建了NAT10敲低的稳定细胞系。进行CCK - 8、EdU检测和集落形成检测以评估细胞体外增殖和克隆形成能力。同时，进行Transwell检测以评估对侵袭和转移能力的影响。通过相关性分析验证NAT10与ZEB1表达之间的相关性。通过qPCR、蛋白质免疫印迹、RIP - qPCR、点图和mRNA稳定性检测证实NAT10调节ZEB1的潜在机制。此外，使用类似的实验方法证实NAT10、ZEB1和MT1 - MMP之间的相互作用。涉及ZEB1过表达的挽救实验进一步验证了NAT10/ZEB1/MT1 - MMP轴在PC转移中的作用。此外，NAT10抑制剂Remodelin用于裸鼠原位PC模型，以研究其对体内转移的影响。

结果

发现NAT10在PC中上调，且与预后不良显著相关。NAT10敲低后，AsPC - 1和KPC的增殖和转移能力明显受损，ac4C修饰程度降低，ZEB1的mRNA稳定性下降。相关性分析表明NAT10、ZEB1和MT1 - MMP之间呈正相关，qPCR和蛋白质免疫印迹结果也证实了这一结论。此外，ZEB1过表达可显著逆转AsPC - 1中NAT10缺失诱导的迁移和侵袭抑制。NAT10抑制剂Remodelin治疗可降低体内腹膜和肝脏转移程度。