TGF-β/Smad3 stimulates stem cell/developmental gene expression and vascular smooth muscle cell de-differentiation.
TGF-β/Smad3 stimulates stem cell/developmental gene expression and vascular smooth muscle cell de-differentiation.
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DOI：
10.1371/journal.pone.0093995
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Kent KC
中科院分区：
文献类型：
--
作者：
Shi X;DiRenzo D;Guo LW;Franco SR;Wang B;Seedial S;Kent KC
动脉粥样硬化相关疾病是美国的主要死亡原因。尽管最近取得了进展，动脉粥样硬化的介入治疗可能会因新内膜增生导致的再狭窄而复杂化。我们以前的研究表明，TGF-β及其下游信号蛋白Smad 3 ∶1）在血管损伤后上调，2）共同驱动平滑肌细胞（SMC）增殖和迁移，3）促进内膜增生的发展。为了确定TGF-β/Smad 3促进这些作用的机制，在用Smad 3感染并用TGF-β刺激或单独用GFP感染的原代大鼠SMC上进行Affyptase基因表达阵列。超过200个基因在TGF-β/Smad 3刺激的SMC中差异表达（>2.0倍变化，p<0.05）。然后，我们使用大卫生物信息学数据库进行了GO术语富集分析，发现TGF-β/Smad 3激活了与发育或细胞分化相关的多个基因的表达，其中一些基因已被证明与多能干细胞或祖细胞相关。定量实时PCR证实了几个发育基因的上调，包括FGF 1，NGF和Wnt 11（分别为2.5，6和7倍）以及干/祖细胞相关基因CD 34和CXCR 4（分别为10和45倍）。此外，蛋白质印迹法或免疫细胞化学（针对CXCR 4和NGF进行）也证实了这些因子在蛋白质水平上的上调。最后，TGF-β/Smad 3下调SMC收缩基因的转录以及平滑肌α肌动蛋白、钙调蛋白和平滑肌肌球蛋白重链的蛋白质产生。这些综合结果表明，TGF-β/Smad 3刺激通过上调发育相关基因驱动SMC进入表型改变的去分化状态。
Atherosclerotic-associated diseases are the leading cause of death in the United States. Despite recent progress, interventional treatments for atherosclerosis can be complicated by restenosis resulting from neo-intimal hyperplasia. We have previously demonstrated that TGF-β and its downstream signaling protein Smad3∶1) are up-regulated following vascular injury, 2) together drive smooth muscle cell (SMC) proliferation and migration and 3) enhance the development of intimal hyperplasia. In order to determine a mechanism through which TGF-β/Smad3 promote these effects, Affymetrix gene expression arrays were performed on primary rat SMCs infected with Smad3 and stimulated with TGF-β or infected with GFP alone. More than 200 genes were differentially expressed (>2.0 fold change, p<0.05) in TGF-β/Smad3 stimulated SMCs. We then performed GO term enrichment analysis using the DAVID bioinformatics database and found that TGF-β/Smad3 activated the expression of multiple genes related to either development or cell differentiation, several of which have been shown to be associated with multipotent stem or progenitor cells. Quantitative real-time PCR confirmed up-regulation of several developmental genes including FGF1, NGF, and Wnt11 (by 2.5, 6 and 7 fold, respectively) as well as stem/progenitor cell associated genes CD34 and CXCR4 (by 10 and 45 fold, respectively). In addition, up-regulation of these factors at protein levels were also confirmed by Western blotting, or by immunocytochemistry (performed for CXCR4 and NGF). Finally, TGF-β/Smad3 down regulated transcription of SMC contractile genes as well as protein production of smooth muscle alpha actin, calponin, and smooth muscle myosin heavy chain. These combined results suggest that TGF-β/Smad3 stimulation drives SMCs to a phenotypically altered state of de-differentiation through the up-regulation of developmental related genes.
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影响因子：
5.3
作者：
Ferlosio, Amedeo;Arcuri, Gaetano;Orlandi, Augusto
通讯作者：
Orlandi, Augusto
影响因子：
4.6
作者：
Cohen, Ethan David;Miller, Mayumi F.;Morrisey, Edward E.
通讯作者：
Morrisey, Edward E.
影响因子：
2.2
作者：
Hollenbeck, ST;Sakakibara, K;Kent, KC
通讯作者：
Kent, KC
影响因子：
5.3
作者：
Imamura, Hirotoshi;Ohta, Tsuyoshi;Kikuta, Ken-ichiro
通讯作者：
Kikuta, Ken-ichiro
影响因子：
2.6
作者：
Crcareva, A;Saito, T;Chiba, S
通讯作者：
Chiba, S