Nox4 involvement in TGF-beta and SMAD3-driven induction of the epithelial-to-mesenchymal transition and migration of breast epithelial cells.
Nox4 involvement in TGF-beta and SMAD3-driven induction of the epithelial-to-mesenchymal transition and migration of breast epithelial cells.
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DOI：
10.1016/j.freeradbiomed.2012.06.016
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发表时间：
2012-10-01
影响因子：
7.4
通讯作者：
Leto, Thomas L.
中科院分区：
文献类型：
--
作者：
Boudreau, Howard E.;Casterline, Benjamin W.;Rada, Balazs;Korzeniowska, Agnieszka;Leto, Thomas L.
关键词：
上皮细胞向间充质细胞转化（EMT）是在伤口愈合和胚胎发育过程中正常发生的细胞可塑性增加的发展，并且可以用于癌症侵袭和转移。TGF-β诱导EMT，但其机制尚不清楚。我们的研究表明，Nox 4，NADPH氧化酶（Nox）家族的成员，是一个来源的活性氧（ROS）影响细胞迁移和纤连蛋白的表达，EMT标记，在正常和转移性乳腺上皮细胞。我们发现TGF-β诱导正常（MCF 10A）和转移性（MDA-MB-231）人乳腺上皮细胞中的Nox 4表达（mRNA和蛋白质）和ROS生成。相反，表达显性阴性形式的Nox 4或Nox 4靶向shRNA的细胞在TGF-β处理后显示出显著较低的ROS产生。组成型活性TGF-β受体I型的表达显著增加了Nox 4启动子活性、mRNA和蛋白质表达以及ROS生成。基于组成型活性SMAD 3增加Nox 4启动子活性的作用，TGF-β对Nox 4转录的调节是SMAD 3依赖性的，而显性负性SMAD 3或SIS 3（SMAD 3特异性抑制剂）具有相反的作用。此外，Nox 4敲低、显性负性Nox 4或SMAD 3或SIS 3钝化TGF-β诱导的伤口愈合和细胞迁移，而细胞增殖不受影响。基于显性负性Nox 4在TGF-β处理的MDA-MB-231和MCF 10A细胞中降低纤连蛋白mRNA的作用，我们的实验进一步表明Nox 4在TGF-β调节纤连蛋白mRNA表达中起作用。总的来说，这些数据表明Nox 4有助于NADPH氧化酶依赖性ROS的产生，这可能是乳腺上皮细胞中EMT进展的关键，从而具有治疗意义。
The epithelial-to-mesenchymal transition (EMT) is the development of increased cell plasticity that occurs normally during wound healing and embryonic development and can be coopted for cancer invasion and metastasis. TGF-beta induces EMT but the mechanism is unclear. Our studies suggest Nox4, a member of the NADPH oxidase (Nox) family, is a source of reactive oxygen species (ROS) affecting cell migration and fibronectin expression, an EMT marker, in normal and metastatic breast epithelial cells. We found TGF-beta induces Nox4 expression (mRNA and protein) and ROS generation in normal (MCF10A) and metastatic (MDA-MB-231) human breast epithelial cells. Conversely, cells expressing a dominant-negative form of Nox4 or Nox4-targeted shRNA showed significantly lower ROS production upon TGF-beta treatment. Expression of a constitutively active TGF-beta receptor type I significantly increased Nox4 promoter activity, mRNA and protein expression, and ROS generation. Nox4 transcriptional regulation by TGF-beta was SMAD3-dependent based on the effect of constitutively active SMAD3 increasing Nox4 promoter activity, whereas dominant-negative SMAD3 or SIS3, a SMAD3-specific inhibitor, had the opposite effect. Furthermore, Nox4 knockdown, dominant-negative Nox4 or SMAD3, or SIS3 blunted TGF-beta induced wound healing and cell migration, whereas cell proliferation was not effected. Our experiments further indicate Nox4 plays a role in TGF-beta regulation of fibronectin mRNA expression, based on the effects of dominant-negative Nox4 in reducing fibronectin mRNA in TGF-beta treated MDA-MB-231and MCF10A cells. Collectively, these data indicate Nox4 contributes to NADPH oxidase-dependent ROS production that may be critical for progression of the EMT in breast epithelial cells, and thereby has therapeutic implications.
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影响因子：
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作者：
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通讯作者：
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影响因子：
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