Hepatocyte Growth Factor Activator Inhibitor Type 1 Regulates Epithelial to Mesenchymal Transition through Membrane-Bound Serine Proteinases
Hepatocyte Growth Factor Activator Inhibitor Type 1 Regulates Epithelial to Mesenchymal Transition through Membrane-Bound Serine Proteinases
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DOI：
10.1158/0008-5472.can-08-3728
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发表时间：
2009-03-01
期刊：
影响因子：
11.2
通讯作者：
Kataoka, Hiroaki
中科院分区：
文献类型：
--
作者：
Cheng, Haixia;Fukushima, Tsuyoshi;Kataoka, Hiroaki
肝细胞生长因子激活物抑制物-1(HAI-1)由丝氨酸蛋白酶抑制因子1(SPLVT1)基因编码，是一种膜相关的蛋白水解酶抑制物，能有效抑制多种丝氨酸蛋白酶，包括膜结合的丝氨酸蛋白酶。尽管HAI-1/SPINT1在上皮细胞和癌细胞中广泛表达，但其功能作用仍不清楚，尤其是在癌症中。在这里，我们展示了在人胰腺癌细胞系Suit-2中稳定地敲除HAI-1/SPINT1诱导了与加速侵袭相关的细长的纺锤形形态，从而模拟了上皮向间充质的转变(EMT)。我们发现，HAI-1/SPINT1基因敲除显著降低了E-钙粘蛋白的表达，并伴随着E-钙粘蛋白转录抑制因子Smad相互作用蛋白1(SUP1)的上调。此外，基质金属蛋白酶-9(MMP9)表达上调。在HLC-1肺癌细胞系中也得到了类似的结果。此外，显示E-eherin表达缺失的Suit-2转移变异体(S2-CP8)也显示HAI-1/SPINT1水平显著降低。基因工程HAI-1/SPINT1在S2-CP8中的过表达导致E-钙粘蛋白表达逆转和SIPI下调，并伴随着培养上皮形态的重建。HAI-1/SPINT1基因敲除的SLTIT-2细胞和HLC-1细胞中的丝氨酸酶/ST14和TMPRSS4基因敲除后，SIP1和/或MMP9的表达水平发生逆转，HAI-1/SPINT1基因敲除引起的EMT可能至少部分由膜结合的丝氨酸酶/ST14和TMPRSS4介导。我们认为，HAI-1/SPINT1和膜结合丝氨酸蛋白酶之间的相互作用有助于某些癌细胞中参与EMT的转录和功能变化。[癌症资源2009；69(5)：1828-35]
Hepatocyte growth factor activator inhibitor-1 (HAI-1), encoded by the serine protease inhibitor Kunitz type 1 (SPLVT1) gene, is a membrane-associated proteinase inhibitor that potently inhibits a variety of serine proteinases, including those that are membrane bound. Although HAI-1/SPINT1 is widely expressed by epithelial cells and cancer cells, its functional role is still unclear, particularly in cancer. Here, we show that stable knockdown of HAI-1/SPINT1 in the human pancreatic cancer cell line SUIT-2 induces an elongated spindle-like morphology associated with accelerated invasion, thereby mimicking an epithelial to mesenchymal transition (EMT). We found that HAI-1/SPINT1 knockdown significantly reduced the expression of E-cadherin and was accompanied by up-regulation of Smad-interacting protein 1 (SUP1), an E-cadherin transcriptional repressor. In addition, matrix metalloproteinase-9 (MMP-9) was up-regulated. Similar results were obtained in the HLC-1 lung carcinoma cell line. Moreover, a metastatic variant of SUIT-2 (S2-CP8) that showed loss of E-eadherin expression also showed a significantly reduced level of HAI-1/SPINT1. Engineered overexpression of HAI-1/SPINT1 in S2-CP8 resulted in reversion of E-cadherin expression and SIPI down-regulation, which accompanied reestablishment of epithelial morphology in culture. The EMT caused by HAI-1/SPINT1 knockdown seemed to be mediated at least partly, by membrane-bound serine proteinases: matriptase/ST14 and TMPRSS4, as knockdown of matriptase/ST14 or TMPRSS4 in HAI-1/SPINT1 knockdown SLTIT-2 cells and HLC-1 cells resulted in reversion of SIP1 and/or MMP-9 expression levels. We suggest that interactions between HAI-1/SPINT1 and membrane-bound serine proteinases contribute to transcriptional and functional changes involved in EMT in certain carcinoma cells. [Cancer Res 2009;69(5):1828-35]