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

IF 3.2 3区医学 Q2 ONCOLOGY Clinical & Experimental Metastasis Pub Date : 2025-01-22 DOI:10.1007/s10585-024-10323-0

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

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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.

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m1a调控的DIAPH3通过稳定KRT19促进结直肠癌的侵袭性。

背景：近年来，由于人们对n1 -甲基腺苷（m1A）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，研究重点已转移到了解m1A在肿瘤进展中的作用。方法：我们对3例结直肠癌患者的肿瘤组织和肿瘤邻近正常组织进行甲基化RNA免疫沉淀测序，以确定结直肠癌中mRNA的m1A谱。利用免疫组织化学和在线数据集评估透明膜相关双胍3 （DIAPH3）的表达及其与CRC临床病理特征的相关性。采用创面愈合实验、Transwell实验和异种移植转移模型来评估膜片3在结直肠癌细胞迁移和侵袭中的作用。利用质谱法筛选了膜片3的下游靶点。通过将DIAPH3 siRNA与角蛋白19 （keratin 19, KRT19）异位质粒共转染CRC细胞，证实了DIAPH3-KRT19信号轴的作用。结果：在结直肠癌组织中，膜片3及其m1A修饰的mRNA水平同时升高。此外，在结直肠癌组织中，高表达的DIAPH3与转移和进展到晚期显著相关。在敲低DIAPH3后，CRC细胞的迁移和侵袭能力明显下降，这可以通过过表达KRT19来挽救。此外，蛋白酶体抑制剂MG132可以阻断由DIAPH3沉默诱导的KRT19降解。结论：我们的研究表明，在CRC细胞中，通过m1A甲基化修饰了diaph3mrna。沉默DIAPH3抑制CRC细胞的迁移和侵袭，可能是通过蛋白酶体依赖下游KRT19的降解。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Clinical & Experimental Metastasis 医学-肿瘤学

CiteScore

7.80

自引率

5.00%

发文量

审稿时长

12 months

期刊介绍： The Journal''s scope encompasses all aspects of metastasis research, whether laboratory-based, experimental or clinical and therapeutic. It covers such areas as molecular biology, pharmacology, tumor biology, and clinical cancer treatment (with all its subdivisions of surgery, chemotherapy and radio-therapy as well as pathology and epidemiology) insofar as these disciplines are concerned with the Journal''s core subject of metastasis formation, prevention and treatment.