A novel IL-10 signalling mechanism regulates TIMP-1 expression in human prostate tumour cells.
A novel IL-10 signalling mechanism regulates TIMP-1 expression in human prostate tumour cells.
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一种新的 IL-10 信号传导机制调节人前列腺肿瘤细胞中的 TIMP-1 表达。
DOI：
10.1038/sj.bjc.6600855
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发表时间：
2003
影响因子：
8.8
通讯作者：
Stearns,ME
中科院分区：
文献类型：
--
作者：
Wang,M;Hu,Y;Stearns,ME
我们先前已经报道，白介素10(IL-10)信号刺激称为hTE-1的特定增强子元件的激活，以促进人骨转移PC-3亚克隆(PC-3 ML)细胞中基质金属蛋白酶组织抑制因子-1(TIMP-1)的表达。最近，我们发现了一个IL-10应答信号分子，命名为IL-10E1，它与hte-1元件结合，并克隆了编码22 kDa蛋白的基因。在这篇论文中，我们研究了两个不同的人前列腺细胞系IL-10/IL-10受体信号转导的机制，一个是名为NPTX-1532的正常前列腺上皮细胞系，另一个是高转移的PC-3ML肿瘤细胞。信号级联研究表明，IL-10刺激JAK1和TYK2受体激酶的酪氨酸磷酸化和IL-10E1的酪氨酸磷酸化。磷酸化，触发IL-10E_1‘S在10-30分钟内快速易位到细胞核。缺失分析和瞬时转染实验表明，IL-10刺激NT-∼多肽的N-末端结构域(TIMP-1)转位到细胞核，诱导TIMP-1表达。定点突变进一步表明，NT-NLS的两个酪氨酸部分(Y57和Y62)的磷酸化是激活IL-10信号和TIMP-1表达所必需的。这些数据首次证明，在人类前列腺细胞中，IL-10受体对TIMP-1表达的信号是通过新基因IL-10E1的酪氨酸磷酸化来调节的。
We have previously reported that interleukin 10 (IL-10) signalling stimulated activation of a specific enhancer element, termed HTE-1, to promote tissue inhibitor of matrix metalloproteinase1 (TIMP-1) expression in human bone metastatic PC-3 subclone (PC-3 ML) cells. Recently, we have identified an IL-10 responsive signal molecule, termed IL-10E1, which binds the HTE-1 element and cloned the gene encoding for the 22 kDa protein. In this paper, we have examined the mechanism of IL-10/IL-10 receptor signalling in two distinct human prostate cell lines, a ‘normal’prostate epithelial cell line, termed NPTX-1532 and highly metastatic PC-3 ML tumour cells. Signalling cascade studies revealed that IL-10 stimulated tyrosine phosphorylation of JAK1 and TYK2 receptor kinases and tyrosine phosphorylation of IL-10E1. Phosphorylation, triggered IL-10E1's rapid translocation to the nucleus by 10–30 min. Deletion analysis combined with transient transfection experiments revealed that the n-terminal domain (∼ 74 aa) of the IL-10E1 protein, the nt-nls peptide, was stimulated by IL-10 to translocate to the nucleus and induce TIMP-1 expression. Site-directed mutagenesis further showed that phosphorylation of two tyrosine moieties (Y57 and Y62) of the nt-nls peptide was required for IL-10 activation of signalling and TIMP-1 expression. The data demonstrate, for the first time, that IL-10 receptor signalling of TIMP-1 expression is regulated by tyrosine phosphorylation of a novel gene, IL-10E1, in human prostate cells.