miR-149-3p–mediated TIMP3 restoration suppresses tumor aggressiveness in sorafenib-resistant liver cancer cell

IF 3.1 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms Pub Date : 2026-03-01 Epub Date: 2026-01-15 DOI:10.1016/j.bbagrm.2026.195136

Sanghee Park , Taehyun Park , Dayoung Choi , Soeun Kim , Hyunjung Cho , Misu Lee

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Abstract

Hepatocellular carcinoma (HCC) is a highly lethal malignancy in which acquired resistance to sorafenib remains a major therapeutic challenge. To investigate mechanisms of resistance, we performed transcriptomic profiling of sorafenib-resistant HCC cells, which revealed enrichment of processes related to tumor progression, including mitotic regulation, chromatin remodeling, and apoptotic signaling. Notably, these cells displayed marked downregulation of tissue inhibitor of metalloproteinases-3 (TIMP3), a tumor suppressor known to regulate invasion and epithelial–mesenchymal transition (EMT). Functional studies showed that TIMP3 overexpression (OV) suppressed cell viability, stemness-associated proteins, and a key cell cycle regulator, while upregulating apoptosis-related proteins. Conversely, TIMP3 knockdown (KD) enhanced proliferation, stemness, and EMT. EMT markers were reduced in TIMP3-OV cells but increased with TIMP3-KD, consistent with spheroid sprouting assays, highlighting TIMP3 as a brake on aggressiveness. To explore upstream regulators, we integrated in silico predictions with validation, identifying miR-149-3p as a novel repressor of TIMP3. miR-149-3p overexpression reduced TIMP3 levels and promoted EMT and invasion, whereas inhibition of miR-149-3p restored TIMP3 expression and suppressed migration. Clinical analyses using TCGA datasets and HCC tissue microarrays confirmed significantly lower TIMP3 expression in tumors compared with normal liver tissue, and showed inverse correlations between TIMP3 and proliferative or stemness markers, including Ki67 and CD44. Collectively, these findings establish a mechanistic axis in which miR-149-3p–mediated suppression of TIMP3 promotes sorafenib resistance, EMT, and stemness in HCC. This work identifies TIMP3 as a pivotal determinant of tumor aggressiveness and suggests restoring TIMP3 or targeting downstream pathways as strategies to overcome resistance.

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mir -149-3p介导的TIMP3恢复抑制索拉非尼耐药肝癌细胞的肿瘤侵袭性。

肝细胞癌（HCC）是一种高度致命的恶性肿瘤，获得性索拉非尼耐药仍然是一个主要的治疗挑战。为了研究耐药机制，我们对索拉非尼耐药的HCC细胞进行了转录组学分析，发现与肿瘤进展相关的过程丰富，包括有丝分裂调节、染色质重塑和凋亡信号传导。值得注意的是，这些细胞显示出金属蛋白酶组织抑制剂-3 （TIMP3）的显著下调，TIMP3是一种已知的调节侵袭和上皮-间质转化（EMT）的肿瘤抑制因子。功能研究表明，TIMP3过表达（OV）抑制细胞活力、干细胞相关蛋白和一个关键的细胞周期调节因子，同时上调凋亡相关蛋白。相反，TIMP3敲低（KD）可增强细胞增殖、干性和EMT。EMT标记物在TIMP3- ov细胞中减少，但在TIMP3- kd细胞中增加，与球形发芽实验一致，突出TIMP3是侵袭性的抑制因素。为了探索上游调节因子，我们整合了计算机预测和验证，确定miR-149-3p是TIMP3的新型抑制因子。过表达miR-149-3p可降低TIMP3水平，促进EMT和侵袭，而抑制miR-149-3p可恢复TIMP3表达，抑制迁移。使用TCGA数据集和HCC组织芯片的临床分析证实，与正常肝组织相比，TIMP3在肿瘤中的表达显著降低，TIMP3与增殖或干性标志物（包括Ki67和CD44）呈负相关。总的来说，这些发现建立了一个机制轴，其中mir -149-3p介导的TIMP3抑制促进了HCC中的索拉非尼耐药、EMT和干细胞。这项工作确定TIMP3是肿瘤侵袭性的关键决定因素，并建议恢复TIMP3或靶向下游途径作为克服耐药性的策略。

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

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms 生物-生化与分子生物学

CiteScore

9.20

自引率

2.10%

发文量

审稿时长

44 days

期刊介绍： BBA Gene Regulatory Mechanisms includes reports that describe novel insights into mechanisms of transcriptional, post-transcriptional and translational gene regulation. Special emphasis is placed on papers that identify epigenetic mechanisms of gene regulation, including chromatin, modification, and remodeling. This section also encompasses mechanistic studies of regulatory proteins and protein complexes; regulatory or mechanistic aspects of RNA processing; regulation of expression by small RNAs; genomic analysis of gene expression patterns; and modeling of gene regulatory pathways. Papers describing gene promoters, enhancers, silencers or other regulatory DNA regions must incorporate significant functions studies.