CCN2 is necessary for the function of mouse embryonic fibroblasts
CCN2 is necessary for the function of mouse embryonic fibroblasts
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DOI：
10.1016/j.yexcr.2006.12.006
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发表时间：
2007-03-10
影响因子：
3.7
通讯作者：
Leask, Andrew
中科院分区：
文献类型：
--
作者：
Kennedy, Laura;Liu, Shangxi;Leask, Andrew
CCN 2由经历活跃组织重塑的间充质细胞表达，并且在结缔组织病理学如纤维化和癌症中特征性地过表达。然而，CCN 2的生理作用和作用机制在很大程度上是未知的。在这里，我们使用全基因组mRNA表达谱、蛋白质组学和功能生物测定分析来探测CCN 2对小鼠胚胎成纤维细胞（MEFs）生物学的贡献。我们表明，ccn 2-/-小鼠胚胎成纤维细胞（MEFs）显着降低了促粘附，促炎症和促血管生成基因，如白细胞介素-6（IL-6），血浆铜蓝蛋白，血小板反应蛋白-1，lipocalin-2和syndecan 4的表达。抗多配体蛋白聚糖4抗体减少ccn 2 +/+ MEF中的ERK磷酸化。在ccn 2 +/+ MEFs中，MEK抑制剂U 0126和显性负性ras降低了IL-6和脂质运载蛋白-2的表达。在ccn 2-/- MEFs中过表达多配体蛋白聚糖4恢复了IL-6和脂质运载蛋白-2 mRNA的表达。Syndecan 4已显示介导细胞迁移。我们发现，ccn 2 +/+ MEFs的迁移速度明显快于ccn 2-/- MEFs;抗多配体蛋白聚糖4抗体和U 0126减少了ccn 2 +/+ MEFs向ccn 2-/- MEFs的迁移。这些结果共同支持多配体蛋白聚糖4在MEF中CCN 2下游起作用的观点，并且减少的多配体蛋白聚糖4表达有助于至少部分的ccn 2-/-表型。此外，这些结果表明，CCN 2是MEFs所必需的，有助于组织重塑的各个方面。与此观点一致，尽管ccn 2 +/+ MEF在细胞周边显示出与迁移表型一致的肌动蛋白应力纤维和粘着斑，但ccn 2-/-MEF显示出减少的粘着斑和肌动蛋白应力纤维，以及降低的跨越胶原凝胶基质抵抗力的能力。总的来说，这些结果表明CCN 2提供成纤维细胞正确参与胚胎发生特征所需的基本的非冗余功能，并进一步表明CCN 2可能在成人伤口愈合，组织修复和纤维化中发挥重要作用。(c)2006年爱思唯尔公司All rights reserved.
CCN2 is expressed by mesenchymal cells undergoing active tissue remodeling, and is characteristically overexpressed in connective tissue pathologies such as fibrosis and cancer. However, the physiological roles and mechanism of action of CCN2 are largely unknown. Here, we probe the contribution of CCN2 to the biology of mouse embryonic fibroblasts (MEFs) using genome-wide mRNA expression profiling, proteomic and functional bioassay analyses. We show that ccn2-/- mouse embryonic fibroblasts (MEFs) have significantly reduced the expression of pro-adhesive, pro-inflammatory and pro-angiogenic genes such as interleukin-6 (IL-6), ceruloplasmin, thrombospondin-1, lipocalin-2 and syndecan 4. Anti-syndecan 4 antibody reduced ERK phosphorylation in ccn2+/+ MEFs. In ccn2+/+ MEFs, the MEK inhibitor U0126 and dominant negative ras reduced expression of IL-6 and lipocalin-2. Overexpressing syndecan 4 in ccn2-/- MEFs restored IL-6 and lipocalin-2 mRNA expression. Syndecan 4 has been shown to mediate cell migration. We found that ccn2+/+ MEFs migrated significantly faster than ccn2-/- MEFs; anti-syndecan 4 antibody and U0126 reduced the migration of ccn2+/+ MEFs to that of ccn2-/- MEFs. These results collectively support the notion that syndecan 4 acts downstream of CCN2 in MEFs, and that reduced syndecan 4 expression contributes to at least part of the ccn2-/- phenotype. Further, these results suggest that CCN2 is required for MEFs to contribute to aspects of tissue remodeling. Consistent with this notion, whereas ccn2+/+ MEFs displayed actin stress fibers and focal adhesions at the cell periphery consistent with a migratory phenotype, ccn2-/- MEFs displayed reduced focal adhesions and actin stress fibers, and a reduced ability to transduce forces across a collagen gel matrix. Collectively, these results suggest that CCN2 supplies essential, non-redundant functions required for fibroblasts to properly participate in features of embryogenesis, and further suggest that CCN2 may play essential roles in adult wound healing, tissue repair and fibrogenesis. (c) 2006 Elsevier Inc. All rights reserved.