Investigating the role of c-Jun N-terminal kinases in the proliferation of Werner syndrome fibroblasts using diaminopyridine inhibitors.
Investigating the role of c-Jun N-terminal kinases in the proliferation of Werner syndrome fibroblasts using diaminopyridine inhibitors.
复制标题
DOI：
10.1186/1752-153x-5-83
复制
发表时间：
2011-12-08
影响因子：
--
通讯作者：
Bagley MC
中科院分区：
文献类型：
--
作者：
Davis T;Dix MC;Rokicki MJ;Brook AJ;Widdowson CS;Kipling D;Bagley MC
来源于早老性沃纳综合征的成纤维细胞显示出减少的复制寿命和“应激”形态，这两者都使用MAP激酶抑制剂SB 203580减轻。然而，这些数据的解释是有问题的，因为虽然SB 203580具有应激激活激酶p38和JNK 1/2作为其优选靶点，但它确实显示出相对较低的总体激酶选择性。几行数据支持p38和JNK 1/2活化在控制细胞增殖以及衰老疾病（包括II型糖尿病，Werner综合征个体易患的疾病）的病理学中的作用，因此使得JNK抑制剂的使用作为可能的治疗剂具有吸引力。因此，我们测试了广泛使用的JNK抑制剂SP 600125对WS细胞的增殖和形态的影响。此外，我们合成和测试了两个最近描述的氨基吡啶为基础的抑制剂。SP 600125处理导致WS细胞增殖停止，并导致衰老样细胞表型，其似乎与JNK 1/2的抑制无关。相比之下，在完全抑制JNK 1/2的浓度下使用更具选择性的氨基吡啶CMPD 60对永生化WS细胞的细胞增殖具有积极作用，但对原代WS成纤维细胞的复制寿命没有影响。此外，CMPD 6 o校正了强制降解WS细胞形态。而氨基吡啶CMPD 6 r对WS细胞作用不大。还发现CMDP 60是MK2的弱抑制剂，这可以部分解释其对WS细胞的作用，因为已知MK2通过HSP 27磷酸化参与调节细胞形态，并且被认为在细胞周期停滞中起作用。这些数据表明，总JNK 1/2活性在WS细胞的增殖控制中不起实质性作用。
Fibroblasts derived from the progeroid Werner syndrome show reduced replicative lifespan and a "stressed" morphology, both alleviated using the MAP kinase inhibitor SB203580. However, interpretation of these data is problematical because although SB203580 has the stress-activated kinases p38 and JNK1/2 as its preferred targets, it does show relatively low overall kinase selectivity. Several lines of data support a role for both p38 and JNK1/2 activation in the control of cellular proliferation and also the pathology of diseases of ageing, including type II diabetes, diseases to which Werner Syndrome individuals are prone, thus making the use of JNK inhibitors attractive as possible therapeutics. We have thus tested the effects of the widely used JNK inhibitor SP600125 on the proliferation and morphology of WS cells. In addition we synthesised and tested two recently described aminopyridine based inhibitors. SP600125 treatment resulted in the cessation of proliferation of WS cells and resulted in a senescent-like cellular phenotype that does not appear to be related to the inhibition of JNK1/2. In contrast, use of the more selective aminopyridine CMPD 6o at concentrations that fully inhibit JNK1/2 had a positive effect on cellular proliferation of immortalised WS cells, but no effect on the replicative lifespan of primary WS fibroblasts. In addition, CMPD 6o corrected the stressed WS cellular morphology. The aminopyridine CMPD 6r, however, had little effect on WS cells. CMDP 6o was also found to be a weak inhibitor of MK2, which may partially explain its effects on WS cells, since MK2 is known to be involved in regulating cellular morphology via HSP27 phosphorylation, and is thought to play a role in cell cycle arrest. These data suggest that total JNK1/2 activity does not play a substantial role in the proliferation control in WS cells.