PI3K p110γ overexpression in idiopathic pulmonary fibrosis lung tissue and fibroblast cells: in vitro effects of its inhibition
PI3K p110γ overexpression in idiopathic pulmonary fibrosis lung tissue and fibroblast cells: in vitro effects of its inhibition
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DOI：
10.1038/labinvest.2013.6
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发表时间：
2013-05-01
影响因子：
5
通讯作者：
Vancheri, Carlo
中科院分区：
文献类型：
--
作者：
Conte, Enrico;Gili, Elisa;Vancheri, Carlo
特发性肺纤维化（IPF）是一种进行性纤维增生性疾病，其分子发病机制尚不清楚。在最近的一篇论文中，我们证明了PI 3 K通路在用TGF-β处理的正常人肺成纤维细胞的增殖和分化成肌成纤维细胞中的关键作用。在这项研究中，我们评估了I类PI 3 K p110亚型在IPF肺组织以及组织来源的成纤维细胞系中的表达。此外，我们研究了选择性抑制p110亚型对IPF成纤维细胞增殖和纤维化活性的体外影响。对正常和IPF肺组织进行IHC。通过Western印迹和流式细胞术分析来评估PI 3 K p110亚型的表达水平。从正常和IPF组织中建立成纤维细胞系，并在体外研究选择性药理学抑制以及小干扰RNA的特异性基因沉默的作用。在正常和IPF组织/组织来源的成纤维细胞之间，未观察到PI 3 K p110 α、β和5亚型的表达存在显着差异，而p110 γ在IPF肺匀浆和离体成纤维细胞系中的表达更高。IPF肺中的肌成纤维细胞和细支气管基底细胞表现出强烈的p110 γ免疫反应性。增生细胞核抗原和细胞周期蛋白D1在纤维化灶细胞中也呈阳性染色。此外，p110 γ药理学抑制和基因沉默均能够显著抑制IPF成纤维细胞的增殖率以及α-SMA表达。我们的数据表明，PI 3 K p110 γ亚型可能在IPF的病因病理学中起重要作用，并可能是一个特定的药理学靶点。实验室调查（2013）93，566-576; doi：10.1038/labinvest.2013.6;在线发表2013年2月25日
Idiopathic pulmonary fibrosis (IPF) is a progressive fibroproliferative disease whose molecular pathogenesis remains unclear. In a recent paper, we demonstrated a key role for the PI3K pathway in both proliferation and differentiation into myofibroblasts of normal human lung fibroblasts treated with TGF-beta. In this research, we assessed the expression of class I PI3K p110 isoforms in IPF lung tissue as well as in tissue-derived fibroblast cell lines. Moreover, we investigated the in vitro effects of the selective inhibition of p110 isoforms on IPF fibroblast proliferation and fibrogenic activity. IHC was performed on normal and IPF lung tissue. Expression levels of PI3K p110 isoforms were evaluated by western blot and flow cytometry analysis. Fibroblast cell lines were established from both normal and IPF tissue and the effects of selective pharmacological inhibition as well as specific gene silencing by small interfering RNAs were studied in vitro. No significant differences between normal and IPF tissue/tissue-derived fibroblasts were observed for the expression of PI3K p110 alpha, beta and,5 isoforms whereas p110 gamma was more greatly expressed in both IPF lung homogenates and ex vivo fibroblast cell lines. Myofibroblasts and bronchiolar basal cells in IPF lungs exhibited strong immunoreactivity for p110 gamma. Positive staining for the markers of proliferation proliferating cell nuclear antigen and cyclin D1 was also shown in cells of fibrolastic foci. Furthermore, both p110 gamma pharmacological inhibition and gene silencing were able to significantly inhibit proliferation rate as well as alpha-SMA expression in IPF fibroblasts. Our data suggest that PI3K p110 gamma isoform may have an important role in the etio-pathology of IPF and can be a specific pharmacological target. Laboratory Investigation (2013) 93, 566-576; doi:10.1038/labinvest.2013.6; published online 25 February 2013