Hepatocyte growth factor activator inhibitor type 1 suppresses metastatic pulmonary colonization of pancreatic carcinoma cells
Hepatocyte growth factor activator inhibitor type 1 suppresses metastatic pulmonary colonization of pancreatic carcinoma cells
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DOI：
10.1111/j.1349-7006.2010.01808.x
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发表时间：
2011-02-01
期刊：
影响因子：
5.7
通讯作者：
Kataoka, Hiroaki
中科院分区：
文献类型：
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作者：
Fukushima, Tsuyoshi;Kawaguchi, Makiko;Kataoka, Hiroaki
肝细胞生长因子激活物抑制物-1(HAI-1)是一种跨膜型蛋白水解酶抑制物，可调节膜结合和细胞外丝氨酸蛋白酶的活性。HAI-1有两个kunitz型抑制域，N-末端kunitz域(Kd1)负责抑制已知的靶蛋白。此前，我们曾报道HAI-1在人胰腺癌细胞株Suit-2中的敲除导致上皮细胞向间充质细胞的转变。为了评估HAI-1在转移中的作用，我们在裸鼠实验性肺转移实验中检测了稳定表达HAI-1短发夹RNA或不稳定表达HAI-1短发夹状RNA的Suit-2细胞的转移能力。肺转移的程度通过组织学检查和直接测定人细胞角蛋白19mRNA的水平来证实。小鼠尾静脉注射Suit-2细胞1周后，注射HAI-1基因敲除Suit-2的小鼠有36%(4/11)观察到明显的转移定植，而对照组小鼠无一例转移。2周后HAI-1基因敲除细胞和对照细胞的转移阳性率分别为80%(4/5)和40%(2/5)，4周后分别为82%(9/11)和45%(5/11)。因此，HAI-1的缺失促进了肺转移。共注射重组Kd1可抑制HAI-1-基因敲除Suit-2细胞1周后的转移。此外，重组Kd1恢复了HAI-1基因敲除Suit-2细胞中E-钙粘附素的水平，并降低了其体外侵袭力。提示HAI-1对Suit-2的肺转移有调节作用，Kd1可能对抑制转移癌细胞的扩散有治疗作用。(《癌症科学》2011；102：407-413)
Hepatocyte growth factor activator inhibitor type 1 (HAI-1) is a transmembrane protease inhibitor that regulates the activities of membrane-bound and extracellular serine proteases. HAI-1 has two Kunitz-type inhibitor domains with the N-terminal Kunitz domain (KD1) responsible for inhibiting known target proteases. Previously, we reported that knockdown of HAI-1 in the human pancreatic carcinoma cell line SUIT-2 resulted in epithelial to mesenchymal transition. To evaluate the role of HAI-1 in metastasis, we examined the metastatic capability of SUIT-2 cells that did or did not stably express HAI-1 short-hairpin RNA in an experimental pulmonary metastasis assay using nude mice. The extent of pulmonary metastasis was verified by histological examination and direct measurement of human cytokeratin 19 mRNA levels. One week after injecting SUIT-2 cells into mouse tail veins, apparent metastatic colonization was observed in 36% (4/11) of mice injected with HAI-1-knockdown SUIT-2, whereas none (0/11) of the control mice were positive for metastasis. After 2 weeks the metastasis positive ratios were 80% (4/5) and 40% (2/5), and after 4 weeks the ratios were 82% (9/11) and 45% (5/11) for HAI-1-knockdown and control SUIT-2 cells, respectively. Thus, loss of HAI-1 promoted pulmonary metastasis. Co-injection of recombinant KD1 abolished metastasis produced by HAI-1-knockdown SUIT-2 cells after 1 week. Moreover, recombinant KD1 restored E-cadherin levels in HAI-1 knockdown SUIT-2 cells and reduced their invasiveness in vitro. These data indicate that HAI-1 regulates pulmonary metastasis of SUIT-2, and KD1 may have therapeutic application for inhibiting metastatic cancer cell spreading. (Cancer Sci 2011; 102: 407-413)