KRT17 Promotes the Activation of HSCs via EMT in Liver Fibrosis.
KRT17 Promotes the Activation of HSCs via EMT in Liver Fibrosis.
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DOI：
10.14218/jcth.2021.00101
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发表时间：
2022-04-28
影响因子：
3.6
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中科院分区：
文献类型：
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尽管肝星状细胞（hsc）的激活在肝纤维化的发展中起着核心作用，但hsc激活的机制尚不清楚。角蛋白17 （Keratin 17, KRT17）是中间丝家族的一员，可调控肿瘤细胞的增殖和迁移。本研究旨在阐明KRT17在造血干细胞激活中的作用以及肝纤维化的机制。采用组织芯片免疫组化技术检测KRT17的表达。采用Western blotting和qRT-PCR方法检测人和小鼠纤维化肝组织中KRT17的表达。用TGF-β1重组蛋白和脂肪细胞分化混合物（adicyte differentiation mixture， MDI）分别处理LX-2细胞，诱导和逆转LX-2细胞活化，探讨KRT17与HSC活化的相关性。此外，通过CCK-8、流式细胞术、Transwell和伤口愈合试验，检测转染krt17过表达质粒或小干扰RNA的LX-2细胞的细胞增殖和迁移能力。最后，通过挽救实验探讨KRT17在HSC活化和上皮-间质转化（EMT）中的作用。KRT17在人和小鼠纤维化肝组织中的表达均高于健康肝组织，且与HSC活化呈正相关。KRT17上调可增强LX-2细胞的增殖、迁移、HSC活化和EMT，而KRT17下调可逆转这些作用。TGF-β1重组蛋白加速KRT17介导的EMT、HSC活化和增殖，TGF-β1抑制剂在体外抵消KRT17的作用。KRT17可能通过TGF-β1信号通路促进肝纤维化中HSC的活化、增殖和EMT的发生，KRT17可能成为治疗肝纤维化的治疗靶点。
Although activation of hepatic stellate cells (HSCs) plays a central role in the development of liver fibrosis, the mechanism underlying the activation of HSCs remains unclear. Keratin 17 (KRT17), a member of the intermediate filament family, can regulate tumor cell proliferation and migration. The current study aimed to elucidate the role of KRT17 in the activation of HSCs and the mechanisms underlying liver fibrosis. The expression of KRT17 was determined using immunohistochemistry in tissue microarray. Western blotting and qRT-PCR assays were used to determine the KRT17 expression in fibrotic liver tissues obtained from human subjects and mice. LX-2 cells were treated with TGF-β1 recombinant protein and adipocyte differentiation mixture (MDI) mix to induce and reverse LX-2 cell activation, respectively, in order to explore the correlation between KRT17 and HSC activation. Additionally, cell proliferation and migration abilities of LX-2 cells transfected with KRT17-overexpressing plasmid or small interfering RNA were determined using CCK-8, flow cytometry, Transwell, and wound healing assays. Finally, rescue assay was used to explore the role of KRT17 in HSC activation and epithelial-mesenchymal transition (EMT). The expression of KRT17 was higher in the human and mouse fibrotic liver tissues than in healthy liver tissues, and it was positively correlated with HSC activation. Upregulated KRT17 enhanced proliferation, migration, HSC activation and EMT in LX-2 cells, while knockdown of KRT17 reversed these effects. TGF-β1 recombinant protein accelerated KRT17-mediated EMT, HSC activation and proliferation, while TGF-β1 inhibitor counteracted the effect of KRT17 in vitro. KRT17 promoted HSC activation, proliferation and EMT in hepatic fibrosis probably via TGF-β1 signaling, and KRT17 might serve as a therapeutic target for the treatment of liver fibrosis.