CCN3/NOV small interfering RNA enhances fibrogenic gene expression in primary hepatic stellate cells and cirrhotic fat storing cell line CFSC
CCN3/NOV small interfering RNA enhances fibrogenic gene expression in primary hepatic stellate cells and cirrhotic fat storing cell line CFSC
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DOI：
10.1007/s12079-011-0141-3
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发表时间：
2012-03-01
影响因子：
4.1
通讯作者：
Weiskirchen, Ralf
中科院分区：
文献类型：
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作者：
Borkham-Kamphorst, Erawan;van Roeyen, Claudia R.;Weiskirchen, Ralf
肾母细胞瘤过表达基因编码CCN家族的基质细胞蛋白（CCN 3/NOV），包括CCN 1（CYR 61）、CCN 2（CTGF）、CCN 4（WISP-1）、CCN 5（WISP-2）和CCN 6（WISP-3）。CCN蛋白参与多种细胞类型中的有丝分裂、粘附、凋亡、细胞外基质产生、生长停滞和迁移的调节。与已知为促纤维化蛋白的CCN 2/CTGF相比，CCN 3/NOV在肝纤维化中的生物学作用仍然不清楚。在这项研究中，我们发现肝星状细胞激活后ccn 3/nov mRNA急剧增加，在完全转分化的肌成纤维细胞中达到峰值水平。在实验性肝纤维化模型中，CCN 3/NOV在mRNA和蛋白水平上显著增加。CCN 3/NOV主要存在于沿着组织损伤和修复区域的非实质细胞中。在胆管结扎模型中，CCN 3/NOV主要定位于沿着汇管区，而四氯化碳的重复应用导致CCN 3/NOV的表达主要在小叶中心区。与CCN 2/CTGF相反，促纤维化细胞因子血小板衍生生长因子-B和-D以及转化生长因子-β抑制CCN 3/NOV表达。在体外，CCN 3/NOV siRNA减弱了脂肪储存细胞系CFSC中的迁移，这与表达CCN 3/NOV的各种类型的细胞迁移到组织损伤和再生区域的体内发现完全一致。CCN 3/NOV的抑制增强了原代大鼠肝星状细胞和CFSC中促纤维化标志物蛋白的表达，如α-平滑肌肌动蛋白、I型胶原、纤连蛋白、CCN 2/CTGF和TIMP-1。我们进一步发现，腺病毒过表达CCN 2/CTGF抑制CCN 3/NOV表达，而CCN 3/NOV的过表达以及靶向siRNA抑制CCN 3/NOV均导致CCN 2/CTGF表达增强。这些结果表明CCN行动的复杂性远远超出了经典的阴/阳相互作用。
Nephroblastoma overexpressed gene encodes a matricellular protein (CCN3/NOV) of the CCN family, comprising CCN1 (CYR61), CCN2 (CTGF), CCN4 (WISP-1), CCN5 (WISP-2), and CCN6 (WISP-3). CCN proteins are involved in the regulation of mitosis, adhesion, apoptosis, extracellular matrix production, growth arrest and migration in multiple cell types. Compared to CCN2/CTGF, known as a profibrotic protein, the biological role of CCN3/NOV in liver fibrosis remains obscure. In this study we showed ccn3/nov mRNA to increase dramatically following hepatic stellate cell activation, reaching peak levels in fully transdifferentiated myofibroblasts. In models of experimental hepatic fibrosis, CCN3/NOV increased significantly at the mRNA and protein levels. CCN3/NOV was found mainly in non-parenchymal cells along the areas of tissue damage and repair. In the bile-duct ligation model, CCN3/NOV was localized mainly along portal tracts, while the repeated application of carbon tetrachloride resulted in CCN3/NOV expression mainly in the centrilobular areas. In contrast to CCN2/CTGF, the profibrotic cytokines platelet-derived growth factor-B and -D as well as transforming growth factor-beta suppressed CCN3/NOV expression. In vitro, CCN3/NOV siRNA attenuated migration in the cirrhotic fat storing cell line CFSC well in line with in vivo findings that various types of cells expressing CCN3/NOV migrate into the area of tissue damage and regeneration. The suppression of CCN3/NOV enhanced expression of profibrotic marker proteins, such as alpha-smooth muscle actin, collagen type I, fibronectin, CCN2/CTGF and TIMP-1 in primary rat hepatic stellate cells and in CFSC. We further found that adenoviral overexpression of CCN2/CTGF suppressed CCN3/NOV expression, while the overexpression of CCN3/NOV as well as the suppression of CCN3/NOV by targeting siRNAs both resulted in enhanced CCN2/CTGF expression. These results indicate the complexity of CCN actions that are far beyond the classic Yin/Yang interplay.