P190B RhoGAP overexpression in the developing mammary epithelium induces TGFβ-dependent fibroblast activation.
P190B RhoGAP overexpression in the developing mammary epithelium induces TGFβ-dependent fibroblast activation.
复制标题
DOI：
10.1371/journal.pone.0065105
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Vargo-Gogola T
中科院分区：
文献类型：
--
作者：
Gillette M;Bray K;Blumenthaler A;Vargo-Gogola T
Rho GTP酶介导基质-上皮相互作用，这对乳腺上皮细胞（MEC）形态发生很重要。细胞外基质（ECM）沉积和重组的增加影响MEC形态发生的Rho GTP酶依赖的方式。尽管已经描述了改变的ECM对MEC形态发生的影响，但MEC如何调节基质沉积还不清楚。以前，我们发现p190 B RhoGAP过表达通过诱导与基质改变相关的过度分支来破坏乳腺形态发生。因此，我们假设MEC过表达p190 B调节旁分泌相互作用，影响成纤维细胞活化。使用在体内形态测定和免疫组化分析和原代细胞培养试验相结合，我们发现，p190 B在MEC过表达激活成纤维细胞，导致增加胶原蛋白，纤连蛋白，层粘连蛋白的生产和胶原交联酶赖氨酰氧化酶的表达升高。在p190 B相关的成纤维细胞中，TGF-β效应器SMAD 2的磷酸化和TGF-β靶基因αSma的表达增加，表明TGF-β信号传导的升高促进了成纤维细胞活化。机械张力和TGF-β协同激活成纤维细胞。有趣的是，与对照MEC相比，p190 B过表达MEC的条件培养基中活性TGF-β升高，并且p190 B过表达MEC在胶原凝胶收缩测定中表现出增加的收缩性。这些数据表明，来自p190 B过表达MEC的旁分泌信号可能激活邻近成纤维细胞中的TGF-β信号。为了支持这一点，将条件培养基从过表达p190 B的MEC转移到野生型成纤维细胞上或将过表达p190 B的MEC与野生型成纤维细胞共培养，与用对照CM处理或与对照MEC共培养的成纤维细胞相比，增加了成纤维细胞中SMAD 2磷酸化和ECM基因的mRNA表达。用TGF-β受体抑制剂处理可阻断ECM基因表达和SMAD 2磷酸化的增加。总之，这些数据表明，乳腺上皮中p190 B过表达通过升高的TGF-β旁分泌信号传导诱导成纤维细胞活化。
Rho GTPases mediate stromal-epithelial interactions that are important for mammary epithelial cell (MEC) morphogenesis. Increased extracellular matrix (ECM) deposition and reorganization affect MEC morphogenesis in a Rho GTPase-dependent manner. Although the effects of altered ECM on MEC morphogenesis have been described, how MECs regulate stromal deposition is not well understood. Previously, we showed that p190B RhoGAP overexpression disrupts mammary gland morphogenesis by inducing hyperbranching in association with stromal alterations. We therefore hypothesized that MEC overexpression of p190B regulates paracrine interactions to impact fibroblast activation. Using a combination of in vivo morphometric and immunohistochemical analyses and primary cell culture assays, we found that p190B overexpression in MECs activates fibroblasts leading to increased collagen, fibronectin, and laminin production and elevated expression of the collagen crosslinking enzyme lysyl oxidase. Phosphorylation of the TGF-β effector SMAD2 and expression of the TGF-β target gene αSma were increased in p190B-associated fibroblasts, suggesting that elevated TGF-β signaling promoted fibroblast activation. Mechanical tension and TGF-β cooperate to activate fibroblasts. Interestingly, active TGF-β was elevated in conditioned medium from p190B overexpressing MECs compared to control MECs, and p190B overexpressing MECs exhibited increased contractility in a collagen gel contraction assay. These data suggest that paracrine signaling from the p190B overexpressing MECs may activate TGF-β signaling in adjacent fibroblasts. In support of this, transfer of conditioned medium from p190B overexpressing MECs onto wildtype fibroblasts or co-culture of p190B overexpressing MECs with wildtype fibroblasts increased SMAD2 phosphorylation and mRNA expression of ECM genes in the fibroblasts when compared to fibroblasts treated with control CM or co-cultured with control MECs. The increased ECM gene expression and SMAD2 phosphorylation were blocked by treatment with a TGF-β receptor inhibitor. Taken together, these data suggest that p190B overexpression in the mammary epithelium induces fibroblast activation via elevated TGF-β paracrine signaling.
登录
查看更多内容
影响因子：
3.7
作者：
Guo F;Carter DE;Leask A
通讯作者：
Leask A
影响因子：
4.8
作者：
Gunther, EJ;Belka, GK;Chodosh, LA
通讯作者：
Chodosh, LA
影响因子：
2.7
作者：
Heckman, Brandy M.;Chakravarty, Geetika;Rosen, Jeffrey M.
通讯作者：
Rosen, Jeffrey M.
影响因子：
3.7
作者：
Nakerakanti SS;Bujor AM;Trojanowska M
通讯作者：
Trojanowska M
DOI：
10.1083/jcb.201102147
发表时间：
2012-02-20
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lu P;Weaver VM;Werb Z
通讯作者：
Werb Z