上海交通大学学报(医学版) ›› 2026, Vol. 46 ›› Issue (8): 1037-1052.doi: 10.3969/j.issn.1674-8115.2026.08.005

• 论著 · 基础研究 • 上一篇    

去泛素化酶USP38通过稳定RBM14促进胃癌进展的机制研究

陈赟琪, 那迪娜·帕尔哈提, 张鹏善(), 黄陈()   

  1. 上海交通大学医学院附属第一人民医院胃肠外科,上海 200080
  • 收稿日期:2026-03-02 接受日期:2026-05-18 出版日期:2026-08-13 发布日期:2026-08-13
  • 通讯作者: 黄 陈,主任医师,博士;电子信箱:richard-hc@hotmail.com
    张鹏善,助理研究员,博士;电子信箱:zhangpengshanxc@163.com
  • 作者简介:第一联系人:陈赟琪负责实验设计、实验操作、数据整理、初稿撰写和作图,那迪娜·帕尔哈提负责实验操作、数据整理、作图和文章修改,张鹏善负责实验方法建立、稿件审阅与修改,黄陈负责对稿件进行最终的审阅和修改。所有作者均阅读并同意最终稿件的提交。
  • 基金资助:
    国家自然科学基金(82203751);国家自然科学基金(82472921);上海交通大学“交大之星”计划医工交叉研究基金(24X010301419)

Deubiquitinase USP38 promotes gastric cancer progression by stabilizing RBM14

Chen Yunqi, Paerhati Nadina, Zhang Pengshan(), Huang Chen()   

  1. Department of Gastrointestinal Surgery, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, China
  • Received:2026-03-02 Accepted:2026-05-18 Online:2026-08-13 Published:2026-08-13
  • Contact: Huang Chen, E-mail: richard-hc@hotmail.com
    Zhang Pengshan, E-mail: zhangpengshanxc@163.com.
  • About author:First author contact:Chen Yunqi was responsible for the experimental design, conducting the experiments, data organization, drafting the initial manuscript, and figure preparation. Nadina Paerhati was responsible for conducting the experiments, data organization, figure preparation, and manuscript revision. Zhang Pengshan was responsible for establishing the experimental methods, manuscript review, and revision. Huang Chen was responsible for the final review and revision of the manuscript. All authors have read the final version of the manuscript and consented to its submission.
  • Supported by:
    National Natural Science Foundation of China(82203751);Shanghai Jiao Tong University "JiaoDa Star" Medical-Engineering Interdisciplinary Research Fund(24X010301419)

摘要:

目的·探讨去泛素化酶泛素特异性肽酶38(ubiquitin-specific peptidase 38,USP38)在胃癌中的表达特征、临床预后价值及其促癌分子机制,并鉴定下游关键靶点核糖核酸结合基序蛋白14(RNA-binding motif protein 14,RBM14)。方法·收集50例在上海交通大学医学院附属第一人民医院胃肠外科接受手术治疗的胃癌患者的肿瘤及癌旁组织标本,利用实时荧光定量PCR和蛋白质印迹法验证USP38在胃癌中的表达,并使用癌症基因组图谱(The Cancer Genome Atlas,TCGA)和基因表达综合数据库(Gene Expression Omnibus,GEO)分析胃癌患者临床特征与USP38表达水平的相关性;通过基因集富集分析(Gene Set Enrichment Analysis,GSEA)、基因本体论(Gene Ontology,GO)及京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)预测USP38相关信号通路。结合文献报道与Pearson相关性分析结果筛选USP38的潜在底物。采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)、5-乙炔基-2'-脱氧尿苷(5-ethynyl-2'- deoxyuridine,EdU)试剂盒、克隆形成及划痕实验评估USP38对AGS、HGC-27胃癌细胞增殖和迁移的影响;利用慢病毒载体包装靶向USP38的短发夹核糖核酸(short hairpin RNA,shRNA)构建稳定敲低细胞株,并通过功能回补(rescue)实验验证USP38-RBM14调控轴的生物学功能。结果·TCGA数据库分析显示,USP38在胃癌组织中显著高表达(P<0.001),其表达水平与患者较短的总生存期(Log-rank P<0.001,HR=1.36)及病理分期(P=0.017)呈正相关,GSEA分析发现USP38高表达组显著富集于G2/M期检查点、早期2因子(early 2 factor,E2F)靶点、核斑点及DNA损伤修复等通路(调整后P值均<0.001),且敲低USP38能够下调增殖及间质标志物的表达。功能实验证实敲低USP38可显著抑制胃癌细胞增殖与迁移(均P<0.05)。Pearson相关性分析显示,USP38与其潜在结合蛋白RBM14的表达呈显著正相关(R=0.22,P<0.001)。进一步研究发现,敲低USP38可降低RBM14蛋白水平,而蛋白酶体抑制剂及放线菌酮(cycloheximide,CHX)实验表明USP38在蛋白质翻译后水平稳定RBM14。功能回补实验显示,过表达RBM14可部分恢复USP38敲低对胃癌细胞的抑制效应,证实USP38通过稳定RBM14蛋白发挥促癌作用。结论·USP38在胃癌中高表达且与患者不良预后相关;敲低USP38能够抑制胃癌细胞增殖和迁移。USP38可通过稳定RBM14蛋白促进胃癌进展。

关键词: 胃癌, 泛素特异性肽酶38, 核糖核酸结合基序蛋白14, 去泛素化酶

Abstract:

Objective ·To investigate the expression characteristics and clinical prognostic value of the deubiquitinating enzyme ubiquitin-specific peptidase 38 (USP38) in gastric cancer, as well as its pro-oncogenic molecular mechanism, and to identify the downstream key target RNA-binding motif protein 14 (RBM14). Methods ·Tumor and adjacent non‑tumor tissue specimens were collected from 50 patients with gastric cancer who underwent surgical treatment at the Department of Gastrointestinal Surgery, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine. Quantitative real-time PCR and Western blotting were used to verify the expression of USP38 in gastric cancer, and data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were used to analyze the correlation between the clinical characteristics of patients with gastric cancer and USP38 expression levels. Gene Set Enrichment Analysis (GSEA), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to predict USP38‑related signaling pathways. Potential substrates of USP38 were screened by combining published literature with the results of Pearson correlation analysis. The effects of USP38 on the proliferation and migration of AGS and HGC‑27 gastric cancer cells were assessed using the cell counting kit‑8 (CCK‑8), 5‑ethynyl‑2'‑deoxyuridine (EdU) assay, colony formation assay, and wound healing assay. A stable knockdown cell line was constructed using lentiviral vectors carrying short hairpin RNA (shRNA) targeting USP38, and functional rescue experiments were performed to validate the biological function of the USP38‑RBM14 regulatory axis. Results ·TCGA data analysis showed that USP38 was significantly highly expressed in gastric cancer tissues (P<0.001), and its expression level was positively correlated with shorter overall survival (log‑rank P<0.001, HR=1.36) and pathological stage (P=0.017). GSEA analysis revealed that the high-USP38-expression group was significantly enriched in the G2/M checkpoint, early 2 factor (E2F) targets, nuclear speckles, and DNA damage repair pathways (all adjusted P value<0.001), and knockdown of USP38 downregulated the expression of proliferation- and mesenchymal-related markers. Functional experiments confirmed that USP38 knockdown significantly inhibited the proliferation and migration of gastric cancer cells (P<0.05). Pearson correlation analysis revealed that the expression levels of USP38 and its potential binding protein RBM14 were significantly positively correlated (R=0.22, P<0.001). Further studies revealed that USP38 knockdown decreased RBM14 protein levels, and experiments with a proteasome inhibitor and cycloheximide (CHX) indicated that USP38 stabilized RBM14 at the post‑translational level. Functional rescue experiments showed that RBM14 overexpression partially reversed the inhibitory effect of USP38 knockdown on gastric cancer cells, confirming that USP38 promoted gastric cancer progression by stabilizing RBM14 protein. Conclusion ·USP38 is highly expressed in gastric cancer and is associated with poor patient prognosis; knocking down USP38 can inhibit the proliferation and migration of gastric cancer cells. USP38 exerts an oncogenic effect by stabilizing the RBM14 protein.

Key words: gastric cancer, ubiquitin-specific peptidase 38 (USP38), RNA-binding motif protein 14 (RBM14), deubiquitinase

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