m1A调控的DIAPH3通过稳定KRT19促进结直肠癌的侵袭性。

m1A-regulated DIAPH3 promotes the invasiveness of colorectal cancer via stabilization of KRT19.

作者信息

Mi Shuyi, Hu Jie, Chen Wenwen, Chen Jingyu, Xu Zhipeng, Xue Meng

机构信息

Department of Gastroenterology, The Second Affiliated Hospital of Zhejiang University School of Medicine, No.88, Jiefang Road, Hangzhou, 310009, Zhejiang, China.

Institute of Gastroenterology, Zhejiang University, Hangzhou, China.

出版信息

Clin Exp Metastasis. 2025 Jan 22;42(2):10. doi: 10.1007/s10585-024-10323-0.

DOI:10.1007/s10585-024-10323-0

PMID:39843730

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11754336/

Abstract

BACKGROUND

In recent years, the emphasis has shifted to understanding the role of N1-methyladenosine (m1A) in tumor progression as little is known about its regulatory effect on mRNA and its role in the metastasis of colorectal cancer (CRC).

METHODS

We performed methylated RNA immunoprecipitation sequencing of tumor tissues and tumor-adjacent normal tissues from three patients with CRC to determine the m1A profile of mRNA in CRC. The expression of diaphanous-related formin 3 (DIAPH3) and its correlation with clinicopathological characteristics of CRC were evaluated using immunohistochemistry and online datasets. The role of DIAPH3 in the migration and invasion of CRC cells was evaluated using wound healing assay, Transwell assay and xenograft metastatic model. The downstream targets of DIAPH3 were screened using mass spectrometry. By co-transfecting DIAPH3 siRNA and a keratin 19 (KRT19) ectopic plasmid into CRC cells, the role of DIAPH3-KRT19 signaling axis was confirmed.

RESULTS

The mRNA level of DIAPH3 and its m1A modifications increased simultaneously in the CRC tissues. In addition, high DIAPH3 expression in CRC tissues is significantly associated with metastasis and progression to an advanced stage. After the knockdown of DIAPH3, the migration and invasion capabilities of CRC cells suffered a notable decline, which could be rescued by overexpressing KRT19. In addition, the proteasome inhibitor MG132 could block the degradation of KRT19 induced by DIAPH3 silencing.

CONCLUSIONS

Our study reveals that DIAPH3 mRNA was modified in CRC cells by m1A methylation. Silencing DIAPH3 suppresses the migration and invasion of CRC cells, potentially through the proteasome-dependent degradation of downstream KRT19.

摘要

背景

近年来，人们的重点已转向了解N1-甲基腺苷（m1A）在肿瘤进展中的作用，因为对其对mRNA的调控作用及其在结直肠癌（CRC）转移中的作用知之甚少。

方法

我们对3例CRC患者的肿瘤组织和癌旁正常组织进行了甲基化RNA免疫沉淀测序，以确定CRC中mRNA的m1A图谱。使用免疫组织化学和在线数据集评估透明质酸相关成束蛋白3（DIAPH3）的表达及其与CRC临床病理特征的相关性。使用伤口愈合试验、Transwell试验和异种移植转移模型评估DIAPH3在CRC细胞迁移和侵袭中的作用。使用质谱筛选DIAPH3的下游靶点。通过将DIAPH3 siRNA和角蛋白19（KRT19）异位质粒共转染到CRC细胞中，证实了DIAPH3-KRT19信号轴的作用。