The increase of microRNA-21 during lung fibrosis and its contribution to epithelial-mesenchymal transition in pulmonary epithelial cells.
The increase of microRNA-21 during lung fibrosis and its contribution to epithelial-mesenchymal transition in pulmonary epithelial cells.
复制标题
肺纤维化过程中microRNA-21的增加及其对肺上皮细胞中上皮间质转变的贡献。
DOI：
10.1186/1465-9921-14-95
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发表时间：
2013-09-24
影响因子：
5.8
通讯作者：
Ichinose M
中科院分区：
文献类型：
--
作者：
Yamada M;Kubo H;Ota C;Takahashi T;Tando Y;Suzuki T;Fujino N;Makiguchi T;Takagi K;Suzuki T;Ichinose M
由于组织修复异常而导致成纤维细胞过量且持续积累，导致纤维化疾病，例如特发性肺纤维化。最近的报告揭示了特发性肺纤维化期间 microRNA 的显着变化，并有证据支持 microRNA 在纤维化背景下的肌成纤维细胞分化和上皮间质转化中的作用。据报道，在纤维化过程中，microRNA-21 在肌成纤维细胞中上调，并通过抑制 Smad7 促进转化生长因子-β 信号传导。然而，microRNA-21的表达变化以及microRNA-21在肺纤维化过程中上皮-间质转化中的作用尚未明确。对经盐水或博来霉素治疗的 C57BL/6 J 小鼠的肺和特发性肺纤维化患者的肺标本进行了分析。进行酶消化以分离单个肺细胞。通过流式细胞术细胞分选分离肺上皮细胞。使用定量PCR和原位杂交分析microRNA-21的表达。为了在培养物中诱导上皮-间质转化，将分离的小鼠肺泡II型细胞在存在转化生长因子-β的情况下在纤连蛋白包被的室载玻片上培养，从而产生增强上皮-间质转化的条件。为了研究 microRNA-21 在上皮间质转化中的作用，我们用 microRNA-21 抑制剂转染细胞。从新鲜分离和培养的细胞中分离总RNA。使用定量 PCR 对 MicroRNA-21 以及作为肺泡上皮细胞或间质细胞分化标志物的基因 mRNA 进行定量。从博莱霉素诱导的肺纤维化模型系统中分离的肺上皮细胞，上皮标记基因的表达降低，而间质标记基因的表达增加。在博莱霉素诱导的肺纤维化和人特发性肺纤维化过程中，分离的肺上皮细胞中的 MicroRNA-21 显着上调。在增强上皮-间质转化的条件下，培养的肺泡上皮细胞中的 MicroRNA-21 也上调。在诱导上皮-间质转化的培养条件下，外源施用 microRNA-21 抑制剂可阻止培养的原代小鼠 II 型肺泡细胞中波形蛋白和 α-平滑肌肌动蛋白表达的增加。我们的实验表明，肺纤维化过程中肺上皮细胞中的 microRNA-21 增加，并促进上皮-间质转化。
The excess and persistent accumulation of fibroblasts due to aberrant tissue repair results in fibrotic diseases such as idiopathic pulmonary fibrosis. Recent reports have revealed significant changes in microRNAs during idiopathic pulmonary fibrosis and evidence in support of a role for microRNAs in myofibroblast differentiation and the epithelial-mesenchymal transition in the context of fibrosis. It has been reported that microRNA-21 is up-regulated in myofibroblasts during fibrosis and promotes transforming growth factor-beta signaling by inhibiting Smad7. However, expression changes in microRNA-21 and the role of microRNA-21 in epithelial-mesenchymal transition during lung fibrosis have not yet been defined. Lungs from saline- or bleomycin-treated C57BL/6 J mice and lung specimens from patients with idiopathic pulmonary fibrosis were analyzed. Enzymatic digestions were performed to isolate single lung cells. Lung epithelial cells were isolated by flow cytometric cell sorting. The expression of microRNA-21 was analyzed using both quantitative PCR and in situ hybridization. To induce epithelial-mesenchymal transition in culture, isolated mouse lung alveolar type II cells were cultured on fibronectin-coated chamber slides in the presence of transforming growth factor-β, thus generating conditions that enhance epithelial-mesenchymal transition. To investigate the role of microRNA-21 in epithelial-mesenchymal transition, we transfected cells with a microRNA-21 inhibitor. Total RNA was isolated from the freshly isolated and cultured cells. MicroRNA-21, as well as mRNAs of genes that are markers of alveolar epithelial or mesenchymal cell differentiation, were quantified using quantitative PCR. The lung epithelial cells isolated from the bleomycin-induced lung fibrosis model system had decreased expression of epithelial marker genes, whereas the expression of mesenchymal marker genes was increased. MicroRNA-21 was significantly upregulated in isolated lung epithelial cells during bleomycin-induced lung fibrosis and human idiopathic pulmonary fibrosis. MicroRNA-21 was also upregulated in the cultured alveolar epithelial cells under the conditions that enhance epithelial-mesenchymal transition. Exogenous administration of a microRNA-21 inhibitor prevented the increased expression of vimentin and alpha-smooth muscle actin in cultured primary mouse alveolar type II cells under culture conditions that induce epithelial-mesenchymal transition. Our experiments demonstrate that microRNA-21 is increased in lung epithelial cells during lung fibrosis and that it promotes epithelial-mesenchymal transition.
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DOI：
10.1152/ajplung.00075.2007
发表时间：
2007-12-01
影响因子：
4.9
作者：
He, Mei;Kubo, Hiroshi;Yamaya, Mutsuo
通讯作者：
Yamaya, Mutsuo
影响因子：
3.7
作者：
Steinman RA;Robinson AR;Feghali-Bostwick CA
通讯作者：
Feghali-Bostwick CA
影响因子：
11.2
作者：
Hurteau, Gregory J.;Carlson, J. Andrew;Brock, Graham J.
通讯作者：
Brock, Graham J.
影响因子：
4.5
作者：
Christoffersen, Nanna Ronbjerg;Silahtaroglu, Asli;Lund, Anders H.
通讯作者：
Lund, Anders H.
影响因子：
82.9
作者：
Zeisberg, M;Hanai, J;Kalluri, R
通讯作者：
Kalluri, R