Down-regulation of USP13 mediates phenotype transformation of fibroblasts in idiopathic pulmonary fibrosis.
Down-regulation of USP13 mediates phenotype transformation of fibroblasts in idiopathic pulmonary fibrosis.
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USP13的下调介导特发性肺纤维化成纤维细胞的表型转化
DOI：
10.1186/s12931-015-0286-3
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发表时间：
2015-10-09
影响因子：
5.8
通讯作者：
Dai H
中科院分区：
文献类型：
--
作者：
Geng J;Huang X;Li Y;Xu X;Li S;Jiang D;Liang J;Jiang D;Wang C;Dai H
特发性肺纤维化（IPF）是一种以肺实质成纤维细胞灶和进行性瘢痕形成为特征的致死性疾病。IPF成纤维细胞表现出增殖增加以及迁移和侵袭增强，类似于癌细胞。这种转化样表型的成纤维细胞在肺纤维化的发展中起着重要的作用，但这一机制还没有得到很好的理解。MethodsIn这项研究中，我们比较了基因表达谱在纤维化肺组织从IPF患者和正常肺组织从原发性自发性气胸患者使用的cDNA微阵列研究IPF的发病机制。在cDNA微阵列中，我们发现USP 13在IPF患者的肺组织中减少，这通过免疫组织化学和蛋白质印迹分析的结果进一步证实。然后，我们在MRC-5细胞中使用RNA干扰来抑制USP 13，并通过Western印迹、实时RT-PCR、溴脱氧尿苷掺入和transwell测定来评估其效果。我们还通过免疫共沉淀和免疫荧光染色来鉴定USP 13和PTEN在IPF中的相关性。结果USP 13在成纤维细胞灶和原代IPF成纤维细胞系中的表达水平显著降低。USP 13的缺失导致成纤维细胞转化为具有增强的增殖、迁移和侵袭能力的侵袭性表型。此外，USP 13与PTEN相互作用，并介导肺成纤维细胞中的PTEN泛素化和降解。结论USP 13下调介导了PTEN蛋白丢失和成纤维细胞表型改变，从而在IPF发病机制中起着至关重要的作用。
BackgroundIdiopathic pulmonary fibrosis (IPF) is a fatal disease characterized by fibroblastic foci and progressive scarring of the pulmonary parenchyma. IPF fibroblasts display increased proliferation and enhanced migration and invasion, analogous to cancer cells. This transformation-like phenotype of fibroblasts plays an important role in the development of pulmonary fibrosis, but the mechanism for this is not well understood.MethodsIn this study, we compared gene expression profiles in fibrotic lung tissues from IPF patients and normal lung tissues from patients with primary spontaneous pneumothorax using a cDNA microarray to examine the mechanisms involved in the pathogenesis of IPF. In a cDNA microarray, we found that USP13 was decreased in lung tissues from patients with IPF, which was further confirmed by results from immunohistochemistry and western blot assays. Then, we used RNA interference in MRC-5 cells to inhibit USP13 and evaluated its effects by western blot, real-time RT-PCR, bromodeoxyuridine incorporation, and transwell assays. We also used co-immunoprecipitation and immunofluorescence staining to identify the correlation between USP13 and PTEN in IPF.ResultsUSP13 expression levels were markedly reduced in fibroblastic foci and primary IPF fibroblast lines. The depletion of USP13 resulted in the transformation of fibroblasts into an aggressive phenotype with enhanced proliferative, migratory, and invasive capacities. Additionally, USP13 interacted with PTEN and mediated PTEN ubiquitination and degradation in lung fibroblasts.ConclusionsDown-regulation of USP13 mediates PTEN protein loss and fibroblast phenotypic change, and thereby plays a crucial role in IPF pathogenesis.