Transforming growth factor β1 regulates the expression of CCN2 in human keratinocytes via Smad‐ERK signalling
Transforming growth factor β1 regulates the expression of CCN2 in human keratinocytes via Smad‐ERK signalling
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DOI：
10.1111/iwj.12749
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发表时间：
2017-12
影响因子：
3.1
通讯作者：
E. Kiwanuka;J. Junker;E. Eriksson
中科院分区：
文献类型：
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作者：
E. Kiwanuka;J. Junker;E. Eriksson
结缔组织生长因子（CCN 2/CTGF）和转化生长因子β1（TGF-β1）是皮肤伤口愈合的重要调节因子，但关于它们在上皮细胞谱系中的表达仍存在争议。在这里，我们研究了表皮再生过程中角质形成细胞中CCN 2的表达及其受TGF-β1的调节。在愈合的全层猪伤口的表皮中检测到CCN 2。将人角质形成细胞与或不与10 ng/ml TGF-β1一起孵育，并用10 μM SIS 3或20 μM PD 98059阻断信号通路。采用半定量真实的时间PCR检测CCN 2 mRNA的表达，Western blot检测CCN 2、磷酸化ERK 1/2、ERK 1/2、磷酸化Smad 3和Smad 2/3蛋白的表达。CCN 2在体内伤口前缘的新表皮中瞬时表达。在体外，TGF-β1在2小时（7.5 ± 1.9倍mRNA增加和3.0 ± 0.6倍蛋白增加）和12小时（5.4 ± 1.9倍mRNA增加和3.3 ± 0.6倍蛋白增加）诱导CCN 2表达。与抑制SMAD通路相比，抑制丝裂原活化蛋白激酶（MAPK）通路在降低TGF-β1诱导的CCN 2 mRNA和蛋白表达方面更有效。MAPK通路的抑制对SMAD通路的活性具有最小的影响。CCN 2在角质形成细胞中表达以响应组织损伤或TGF-β1。此外，TGF-β1通过ras/MEK/ERK途径诱导角质形成细胞中的CCN 2表达。全面了解CCN 2在角质形成细胞中的表达对于开发用于伤口愈合和皮肤恶性肿瘤的新疗法至关重要。
Connective tissue growth factor (CCN2/CTGF) and transforming growth factor β1 (TGF‐β1) are important regulators of skin wound healing, but controversy remains regarding their expression in epithelial cell lineages. Here, we investigate the expression of CCN2 in keratinocytes during reepithelialisation and its regulation by TGF‐β1. CCN2 was detected in the epidermis of healing full‐thickness porcine wounds. Human keratinocytes were incubated with or without 10 ng/ml TGF‐β1, and signalling pathways were blocked with 10‐μM SIS3 or 20‐μM PD98059. Semi‐quantitative real‐time PCR was used to study CCN2 mRNA expression, and western blot was used to measure CCN2, phosphorylated‐ERK1/2, ERK1/2, phosphorylated‐Smad3 and Smad2/3 proteins. CCN2 was transiently expressed in neoepidermis at the leading edge of the wound in vivo. In vitro, CCN2 expression was induced by TGF‐β1 at 2 hours (7·5 ± 1·9‐fold mRNA increase and 3·0 ± 0·6‐fold protein increase) and 12 hours (5·4 ± 1·9‐fold mRNA increase and 3·3 ± 0·6‐fold protein increase). Compared with inhibiting the SMAD pathway, inhibiting the mitogen‐activated protein kinase (MAPK) pathway was more effective in reducing TGF‐β1‐induced CCN2 mRNA and protein expression. Inhibition of the MAPK pathway had minimal impact on the activity of the SMAD pathway. CCN2 is expressed in keratinocytes in response to tissue injury or TGF‐β1. In addition, TGF‐β1 induces CCN2 expression in keratinocytes through the ras/MEK/ERK pathway. A complete understanding of CCN2 expression in keratinocytes is critical to developing novel therapies for wound healing and cutaneous malignancy.