USP28敲低和小分子抑制剂促进KRT1不稳定并使肝癌细胞对索拉非尼敏感

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research Pub Date : 2025-04-11 DOI:10.1016/j.yexcr.2025.114558

Zilin Li , Yan Wang , Jiahui Hu , Jingyang Du , Huizong Nie , Yiling Xi , Yue Huang , Kexin Wang , Kaixuan Zhang , Qiuran Xu , Liyan Cheng , Dongsheng Huang , Linglan Tu

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引用次数: 0

摘要

背景：肝细胞癌（HCC）是一种重要的恶性肿瘤，通常诊断较晚且预后差。USP28（泛素特异性蛋白酶28）是泛素特异性蛋白酶（USPs）家族中的一种去泛素化酶，在各种生物过程中，特别是在癌症进展中起着关键作用。然而，其在HCC中的功能和分子机制尚不清楚。方法首先利用TCGA数据库分析USP28在HCC组织中相对于正常组织的表达水平。qRT-PCR和Western Blot进一步验证了这一点。为了研究USP28在HCC中的功能，我们在对照和USP28敲低或过表达的HCC细胞中进行了CCK-8实验、克隆形成实验和Transwell实验。为了探索USP28潜在的下游靶点，我们使用了IP-MS分析。通过免疫沉淀和免疫荧光染色证实USP28与KRT1的相互作用。最后，我们利用异位荷瘤小鼠模型评估了USP28对HCC生长和转移的体内影响。结果USP28在HCC组织中的表达明显高于正常组织，其高表达与预后不良相关。功能实验表明，下调USP28表达可有效抑制HCC细胞的增殖、迁移和侵袭，而过表达USP28则相反。机制研究表明，USP28与KRT1相互作用，使KRT1去泛素化，从而维持KRT1的稳定性。进一步研究发现，USP28敲低导致IFITM3表达降低，抑制HCC细胞增殖。此外，在肿瘤移植小鼠模型中，USP28敲低联合索拉非尼可抑制肿瘤生长和转移。结论sour研究证实了USP28通过稳定KRT1表达和促进IFITM3的致癌作用。USP28小分子抑制剂可以抑制肝癌细胞的增殖，增强肝癌细胞系对索拉非尼的敏感性。这为USP28成为缓解索拉非尼耐药的临床新方法提供了理论依据。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

USP28 knockdown and small molecule inhibitors promote KRT1 destabilization and sensitize hepatocellular carcinoma cells to sorafenib

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USP28 knockdown and small molecule inhibitors promote KRT1 destabilization and sensitize hepatocellular carcinoma cells to sorafenib

Background

Hepatocellular carcinoma (HCC) is a significant malignant tumor that is typically diagnosed late and has a poor prognosis. USP28 (Ubiquitin-specific protease 28), a deubiquitinating enzyme within the ubiquitin-specific proteases (USPs) family, plays a pivotal role in various biological processes, especially in cancer progression. However, its functions and molecular mechanisms in HCC are still unknown.

Methods

We first analyzed the expression level of USP28 in HCC tissues relative to normal tissues using TCGA database. This was further validated by qRT-PCR and Western Blot. To investigate the function of USP28 in HCC, CCK-8 assay, clone formation assay and Transwell assay were performed in control and USP28 knockdown or overexpressed HCC cells. To explore potential downstream targets of USP28, we used IP-MS analysis. The interaction between USP28 and KRT1 was confirmed by immunoprecipitation and immunofluorescence staining. Finally, we evaluated the in vivo effects of USP28 on HCC growth and metastasis using a ectopic tumor-bearing mouse model.

Results

The expression of USP28 in HCC tissues was significantly higher than that in normal tissues, and its high expression was associated with poor prognosis. Functional experiments showed that down-regulation of USP28 expression effectively inhibited the proliferation, migration and invasion of HCC cells, while overexpression of USP28 produced the opposite effect. Mechanistic investigations demonstrated that USP28 interacted with KRT1 and exerted deubiquitination on KRT1, thereby maintaining the stability of KRT1. Further studies revealed that USP28 knockdown resulted in decreased IFITM3 expression, which inhibited HCC cell proliferation. In addition, USP28 knockdown combined with sorafenib inhibited tumor growth and metastasis in tumor xenograft mice model.

Conclusions

Our study confirmed the carcinogenic effects of USP28 by stabilizing KRT1 expression and promoting IFITM3. USP28 small molecule inhibitors can inhibit the proliferation of hepatocellular carcinoma cells and enhance the sensitivity of hepatocellular carcinoma cell lines to sorafenib. This provides a theoretical basis for USP28 to be a new clinical method to alleviate sorafenib resistance.

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来源期刊

Experimental cell research 医学-细胞生物学

CiteScore

7.20

自引率

0.00%

发文量

295

审稿时长

30 days

期刊介绍： Our scope includes but is not limited to areas such as: Chromosome biology; Chromatin and epigenetics; DNA repair; Gene regulation; Nuclear import-export; RNA processing; Non-coding RNAs; Organelle biology; The cytoskeleton; Intracellular trafficking; Cell-cell and cell-matrix interactions; Cell motility and migration; Cell proliferation; Cellular differentiation; Signal transduction; Programmed cell death.