ERK2 but not ERK1 mediates HGF-induced motility in non-small cell lung carcinoma cell lines
ERK2 but not ERK1 mediates HGF-induced motility in non-small cell lung carcinoma cell lines
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DOI：
10.1242/jcs.115832
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发表时间：
2013-06-01
影响因子：
4
通讯作者：
Parker, Peter J.
中科院分区：
文献类型：
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作者：
Radtke, Simone;Milanovic, Mina;Parker, Peter J.
受体酪氨酸激酶（RTK）（如c-Met，肝细胞生长因子（HGF）的受体）的异常信号传导与各种肿瘤（包括非小细胞肺癌（NSCLC））的肿瘤发生有关。通过其促迁移特性，c-Met特别涉及肿瘤转移的过程，需要更好地了解潜在的信号通路。c-Met下游的各种参与者已经得到了很好的表征，包括细胞外信号调节激酶（ERK）1和2。在A549肺癌细胞中进行的基于小干扰RNA（siRNA）的高通量伤口愈合筛选中，我们确定了ERK 2而不是ERK 1作为HGF诱导运动的强介质。这一发现在几种NSCLC细胞系以及HeLa细胞中得到证实。细胞迁移中ERK激酶的一种已知底物，粘着斑蛋白桩蛋白，也是筛选中鉴定的命中物之一。我们证明，肝细胞生长因子的刺激结果在时间依赖性的磷酸化桩蛋白丝氨酸126，这一过程可以通过抑制ERK 1/2上游激酶丝裂原活化蛋白激酶/ERK激酶1（MEK 1）或抑制糖原合成酶激酶3（GSK 3）。此外，我们表明，桩蛋白营业额在局灶性粘连增加刺激后，肝细胞生长因子，这种作用是依赖于丝氨酸残基126（GSK 3网站）和130（ERK网站）内桩蛋白。与ERK 2在肺肿瘤细胞模型中对HGF介导的迁移的亚型特异性要求一致，ERK 2而不是ERK 1被证明负责桩蛋白丝氨酸126磷酸化及其在局灶性粘连处的营业额增加。
Aberrant signalling of receptor tyrosine kinases (RTKs), such as c-Met, the receptor for hepatocyte growth factor (HGF), has been implicated in the oncogenesis of various tumours including non-small cell lung carcinoma (NSCLC). Through its pro-migratory properties, c-Met has been implicated specifically in the process of tumour metastasis, demanding a better understanding of the underlying signalling pathways. Various players downstream of c-Met have been well characterised, including the extracellular-signal-regulated kinases (ERKs) 1 and 2. In a small interfering RNA (siRNA)-based high-throughput wound healing screen performed in A549 lung carcinoma cells, we identified ERK2 but not ERK1 as a strong mediator of HGF-induced motility. This finding was confirmed in several NSCLC cell lines as well as in HeLa cells. One known substrate for ERK kinases in cell migration, the focal adhesion protein paxillin, was also one of the hits identified in the screen. We demonstrate that HGF stimulation results in a time-dependent phosphorylation of paxillin on serine 126, a process that can be blocked by inhibition of the ERK1/2 upstream kinase mitogen-activated protein kinase/ERK kinase 1 (MEK1) or inhibition of glycogen synthase kinase 3 (GSK3). Further, we show that paxillin turnover at focal adhesions is increased upon stimulation by HGF, an effect that is dependent on serine residues 126 (GSK3 site) and 130 (ERK site) within paxillin. In line with the isoform-specific requirement of ERK2 for HGF-mediated migration in lung tumour cell models, ERK2 but not ERK1 is shown to be responsible for paxillin serine 126 phosphorylation and its increased turnover at focal adhesions.