PUS7-dependent pseudouridylation of ALKBH3 mRNA inhibits gastric cancer progression

IF 7.9 1区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL Clinical and Translational Medicine Pub Date : 2024-08-23 DOI:10.1002/ctm2.1811

Yongxia Chang, Hao Jin, Yun Cui, Feng Yang, Kanghua Chen, Wenjun Kuang, Chunxiao Huo, Zhangqi Xu, Ya Li, Aifu Lin, Bo Yang, Wei Liu, Shanshan Xie, Tianhua Zhou

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Abstract

Background

RNA pseudouridylation is a critical post-transcriptional modification that influences gene expression and impacts various biological functions. Despite its significance, the role of mRNA pseudouridylation in cancer remains poorly understood. This study investigates the impact of pseudouridine synthase 7 (PUS7)-mediated pseudouridylation of Alpha-ketoglutarate-dependent Dioxygenase alkB Homolog 3 (ALKBH3) mRNA in gastric cancer.

Methods

Immunohistochemistry and Western blotting were used to assess PUS7 protein levels in human gastric cancer tissues. The relationship between PUS7 and gastric cancer progression was examined using 3D colony formation assays and subcutaneous xenograft models. Real-time quantitative PCR (RT-qPCR), Western blotting, and polysome profiling assays were conducted to investigate how PUS7 regulates ALKBH3. A locus-specific pseudouridine (Ψ) detection assay was used to identify Ψ sites on ALKBH3 mRNA.

Results

Our findings indicate a significant reduction of PUS7 in gastric cancer tissues compared to adjacent non-tumour tissues. Functional analyses reveal that PUS7 inhibits gastric cancer cell proliferation and tumour growth via its catalytic activity. Additionally, PUS7 enhances the translation efficiency of ALKBH3 mRNA by modifying the U696 site with pseudouridine, thereby attenuating tumour growth. Importantly, ALKBH3 functions as a tumour suppressor in gastric cancer, with its expression closely correlated with PUS7 levels in tumour tissues.

Conclusions

PUS7-dependent pseudouridylation of ALKBH3 mRNA enhances its translation, thereby suppressing gastric cancer progression. These findings highlight the potential significance of mRNA pseudouridylation in cancer biology and suggest a therapeutic target for gastric cancer.

Highlights

PUS7 enhances the translation efficiency of ALKBH3 through its pseudouridylation activity on ALKBH3 mRNA, thereby inhibiting gastric tumourigenesis.
The expression levels of PUS7 and ALKBH3 are significantly correlated in gastric tumours, which may be potential prognostic predictors and therapeutic targets for patients with gastric cancer.

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PUS7依赖的ALKBH3 mRNA伪酰化抑制胃癌的进展。

背景：RNA 伪核苷酸化是一种关键的转录后修饰，会影响基因表达并影响各种生物功能。尽管其意义重大，但人们对 mRNA 伪尿嘧啶化在癌症中的作用仍然知之甚少。本研究探讨了假尿苷合成酶7（PUS7）介导的α-酮戊二酸依赖性二氧合酶alkB同源物3（ALKBH3）mRNA假尿苷化对胃癌的影响：方法：采用免疫组织化学和 Western 印迹技术评估人胃癌组织中 PUS7 蛋白水平。采用三维集落形成试验和皮下异种移植模型检测了 PUS7 与胃癌进展之间的关系。研究人员还进行了实时定量 PCR（RT-qPCR）、Western 印迹和多聚体图谱分析，以探讨 PUS7 如何调控 ALKBH3。利用位点特异性假尿嘧啶（Ψ）检测试验确定了 ALKBH3 mRNA 上的Ψ位点：结果：我们的研究结果表明，与邻近的非肿瘤组织相比，胃癌组织中的 PUS7 明显减少。功能分析显示，PUS7 通过其催化活性抑制胃癌细胞增殖和肿瘤生长。此外，PUS7 通过用假尿苷修饰 U696 位点来提高 ALKBH3 mRNA 的翻译效率，从而抑制肿瘤生长。重要的是，ALKBH3在胃癌中发挥着肿瘤抑制因子的作用，其表达量与肿瘤组织中的PUS7水平密切相关：结论：PUS7 依赖的 ALKBH3 mRNA 伪尿苷化可增强其翻译，从而抑制胃癌的进展。这些发现凸显了 mRNA 伪酰化在癌症生物学中的潜在意义，并提出了胃癌的治疗靶点：PUS7通过对ALKBH3 mRNA的假苷酸化活性提高了ALKBH3的翻译效率，从而抑制了胃癌的发生。PUS7和ALKBH3在胃癌中的表达水平显著相关，可能是胃癌患者潜在的预后预测指标和治疗靶点。

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 and Translational Medicine Multiple-

CiteScore

15.90

自引率

1.90%

发文量

450

审稿时长

4 weeks

期刊介绍： Clinical and Translational Medicine (CTM) is an international, peer-reviewed, open-access journal dedicated to accelerating the translation of preclinical research into clinical applications and fostering communication between basic and clinical scientists. It highlights the clinical potential and application of various fields including biotechnologies, biomaterials, bioengineering, biomarkers, molecular medicine, omics science, bioinformatics, immunology, molecular imaging, drug discovery, regulation, and health policy. With a focus on the bench-to-bedside approach, CTM prioritizes studies and clinical observations that generate hypotheses relevant to patients and diseases, guiding investigations in cellular and molecular medicine. The journal encourages submissions from clinicians, researchers, policymakers, and industry professionals.