OTUD6B去泛素化PABPC1促进胃癌细胞的增殖与迁移-东南大学学报医学版

OTUD6B去泛素化PABPC1促进胃癌细胞的增殖与迁移

单位：南通大学附属医院胃肠外科, 江苏南通 226001

分类号：R735.2

出版年·卷·期（页码）：2025·44·第三期（345-354）

摘要：

目的: 探讨卵巢肿瘤结构域去泛素化酶6B(OTUD6B)在胃癌(GC)进展中的作用并明确其相关分子机制。方法: 通过TIMER 2.0数据库,分析OTUD6B在泛癌中的差异表达;通过免疫印迹和qRT-PCR方法,分析胃癌组织以及细胞系中OTUD6B的表达差异,并通过Kaplan-Meier 生存曲线分析OTUD6B的表达与胃癌患者预后的相关性;进一步采用免疫组化(IHC)技术检测胃癌及邻近组织中OTUD6B的表达情况。通过慢病毒介导的基因沉默技术在HGC-27、AGS细胞中敲低OTUD6B的表达;通过CCK-8、EDU实验评估OTUD6B对胃癌细胞增殖能力的影响;通过Transwell、划痕实验探究OTUD6B对胃癌细胞迁移能力的影响;通过免疫共沉淀、免疫荧光技术验证OTUD6B与靶标之间的相互作用。结果:OTUD6B在胃癌组织中表达显著上调,且OTUD6B的异常高表达与胃癌患者的总体不良预后相关。体外实验表明,敲低OTUD6B后,胃癌细胞的增殖和迁移能力明显减弱。在机制层面,OTUD6B可以与 polyA结合蛋白胞质1(PABPC1)直接结合并发挥去泛素化酶作用,减少其K48类型泛素化修饰进而稳定PABPC1的蛋白表达水平。回复实验结果表明过表达PABPC1可以部分逆转OTUD6B敲低所产生的对胃癌进展抑制的效应。结论: OTUD6B可通过结合并去泛素化稳定PABPC1,促进胃癌细胞增殖和迁移,表明OTUD6B可能是胃癌进展的潜在治疗靶标。

Objective: To investigate the role of ovarian tumor(OTU) domain protein 6B(OTUD6B) in the progression of gastric cancer(GC) and to clarify its related molecular mechanism. Methods: The differential expression of OTUD6B was analyzed by TIMER 2.0 database. Western blotting and qRT-PCR were used to analyze the expression differences of OTUD6B in gastric cancer tissues and cell lines, and Kaplan-Meier was used to analyze the correlation between OTUD6B expression and prognosis of patients with gastric cancer. The expression of OTUD6B in gastric cancer and adjacent tissues was further detected by immunohistochemistry(IHC). The expression of OTUD6B in HGC-27 and AGS cells was knocked down by lentivirus-mediated gene silencing technique. The effects of OTUD6B on the proliferation of gastric cancer cells were evaluated by CCK-8 and EDU experiments. The effect of OTUD6B on the migration ability of gastric cancer cells was investigated by transwell and scratch tests. The interaction between OTUD6B and the target was verified by co-immunoprecipitation and immunofluorescence techniques. Results: OTUD6B expression was significantly up-regulated in gastric cancer tissues, and high expression of OTUD6B was associated with poor overall survival. In vitro experiments showed that the proliferation and migration ability of gastric cancer cells were significantly weakened after OTUD6B knockdown. Mechanically, OTUD6B binded and deubiquitinated poly A binding protein cytoplasmic 1(PABPC1), reducing the level of K48 ubiquitination modification and finally stabilizing PABPC1. The response experiment showed that overexpression of PABPC1 partially reversed the knockdown effect of OTUD6B. Conclusion: OTUD6B can stabilize PABPC1 through binding and deubiquitination, and promote the proliferation and migration of gastric cancer cells, suggesting that OTUD6B may be a potential therapeutic target for the progression of gastric cancer.

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