Conditioned media from lung cancer cell line A549 and PC9 inactivate pulmonary fibroblasts by regulating protein phosphorylation.
Conditioned media from lung cancer cell line A549 and PC9 inactivate pulmonary fibroblasts by regulating protein phosphorylation.
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来自肺癌细胞系 A549 和 PC9 的条件培养基通过调节蛋白质磷酸化来灭活肺成纤维细胞。
DOI：
10.1016/j.abb.2011.12.012
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Munakata H.
中科院分区：
文献类型：
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作者：
Park AM;Hayakawa S;Honda E;Mine Y;Yoshida K;Munakata H.
肺纤维化是一种破坏性疾病，由过量的细胞外基质沉积导致进行性肺破坏和瘢痕形成。在纤维化疾病的发病机制中，转化生长因子-β（TGF-β）对肌成纤维细胞的活化起着至关重要的作用。由于目前还没有有效的治疗肺纤维化的方法，找到一种有效的抗纤维化药物是一个重要的目标。一种方法可能是通过鉴定抑制肌成纤维细胞的试剂。在目前的研究中，我们检查了从几种类型的细胞中获得的条件培养基表现出肌成纤维细胞失活活性的潜力。肺癌细胞系A549和PC 9的条件培养基被发现具有这种作用，如通过其降低MRC-5细胞中α-平滑肌肌动蛋白表达的能力所示。随后从培养基中纯化了抑制因子，鉴定为5′-脱氧-5 ′-甲硫基腺苷（MTA），并阐明了其作用机制。检测蛋白激酶A和cAMP反应元件结合蛋白（CREB）的活化。MTA抑制TGF-β诱导的丝裂原活化蛋白激酶的活化。此外，功能获得性突变CREB导致肌成纤维细胞失活。这些结果表明，A549和PC 9条件培养基具有使肌成纤维细胞增殖的能力，并且CREB磷酸化在该过程中起核心作用。
Pulmonary fibrosis is a devastating condition resulting from excess extracellular matrix deposition that leads to progressive lung destruction and scarring. In the pathogenesis of fibrotic diseases, activation of myofibroblasts by transforming growth factor-β (TGF-β) plays a crucial role. Since no effective therapy for pulmonary fibrosis is currently recognized, finding an effective antifibrotic agent is an important objective. One approach might be through identification of agents that inactivate myofibroblasts. In the current study we examined the potential of conditioned medium obtained from several types of cells to exhibit myofibroblast inactivating activity. Conditioned media from lung cancer cell lines A549 and PC9 were found to have this action, as shown by its ability to decrease α-smooth muscle actin expression in MRC-5 cells. Subsequently the inhibitory factor was purified from the medium and identified as 5′-deoxy-5′-methylthioadenosine (MTA), and its mechanism of action elucidated. Activation of protein kinase A and cAMP responsive element binding protein (CREB) were detected. MTA inhibited TGF-β-induced mitogen-activated protein kinase activation. Furthermore, the gain-of-function mutant CREB caused inactivation of myofibroblasts. These results show that A549 and PC9 conditioned media have the ability to inactivate myofibroblasts, and that CREB-phosphorylation plays a central role in this process.