5-HT2A receptor induces ERK phosphorylation and proliferation through ADAM-17 tumor necrosis factor-α-converting enzyme (TACE) activation and heparin-bound epidermal growth factor-like growth factor (HB-EGF) shedding in mesangial cells
5-HT2A receptor induces ERK phosphorylation and proliferation through ADAM-17 tumor necrosis factor-α-converting enzyme (TACE) activation and heparin-bound epidermal growth factor-like growth factor (HB-EGF) shedding in mesangial cells
复制标题
DOI：
10.1074/jbc.m512096200
复制
发表时间：
2006-07-28
影响因子：
4.8
通讯作者：
Raymond, John R.
中科院分区：
文献类型：
--
作者：
Gooz, Monika;Gooz, Pal;Raymond, John R.
在这项研究中，我们提出了多条证据支持肝素结合的EGF（表皮生长因子）样生长因子（HB-EGF）和肿瘤坏死因子-α-转化酶（TACE）（ADAM 17）在肾系膜细胞中EGF受体（EGFR）的反式激活、ERK磷酸化和5-HT 2A受体诱导的细胞增殖中的关键作用。5-羟色胺（5-HT）导致TACE快速激活、HB-EGF脱落、EGFR激活、ERK磷酸化和系膜细胞中DNA含量的长期增加。ERK磷酸化通过以下方式减弱：1）中和EGFR抗体和EGFR激酶抑制剂AG 1478，2）中和HB-EGF，但不中和双调蛋白、抗体、肝素或CM 197，3）基质降解金属蛋白酶或TACE小干扰RNA的药理学抑制剂。外源性HB-EGF刺激ERK磷酸化。此外，TACE与HB-EGF共免疫沉淀。针对TACE的小干扰RNA也阻断了5-HT诱导的ERK磷酸化、HB-EGF脱落和DNA含量的增加。总的来说，这项工作支持可以如下描述的通路图：5-HT -> 5-HT 2A受体-> TACE -> HB-EGF脱落-> EGFR -> ERK ->增加的DNA含量。据我们所知，这是TACE首次参与5-HT诱导的EGFR反式激活或培养的天然细胞中G蛋白偶联受体诱导的增殖。
In this study, we present multiple lines of evidence to support a critical role for heparin-boundEGF( epidermal growth factor)like growth factor (HB-EGF) and tumor necrosis factor-alpha-converting enzyme (TACE) (ADAM17) in the transactivation of EGF receptor (EGFR), ERK phosphorylation, and cellular proliferation induced by the 5-HT2A receptor in renal mesangial cells. 5-hydroxy-tryptamine (5-HT) resulted in rapid activation of TACE, HB-EGF shedding, EGFR activation, ERK phosphorylation, and longer term increases in DNA content in mesangial cells. ERK phosphorylation was attenuated by 1) neutralizing EGFR antibodies and the EGFR kinase inhibitor, AG1478, 2) neutralizing HB-EGF, but not amphiregulin, antibodies, heparin, or CM197, and 3) pharmacological inhibitors of matrix-degrading metalloproteinases or TACE small interfering RNA. Exogenously administered HB-EGF stimulated ERK phosphorylation. Additionally, TACE was co-immunoprecipitated with HB-EGF. Small interfering RNA against TACE also blocked 5-HT-induced increases in ERK phosphorylation, HB-EGF shedding, and DNA content. In aggregate, this work supports a pathway map that can be depicted as follows: 5-HT -> 5-HT2A receptor -> TACE -> HB-EGF shedding -> EGFR -> ERK -> increased DNA content. To our knowledge, this is the first time that TACE has been implicated in 5-HT-induced EGFR transactivation or in proliferation induced by a G protein-coupled receptor in native cells in culture.