Kanemaru Ai, Yamamoto Koji, Kawaguchi Makiko, Fukushima Tsuyoshi, Lin Chen-Yong, Johnson Michael D, Camerer Eric, Kataoka Hiroaki
Section of Oncopathology and Regenerative Biology, Department of Pathology, Faculty of Medicine, University of Miyazaki, Japan.
School of Medicine, Lambardi Comprehensive Cancer Centre, Georgetown University, Washington, DC.
Int J Cancer. 2017 Jan 1;140(1):130-141. doi: 10.1002/ijc.30426. Epub 2016 Sep 23.
Cancer-associated fibroblasts (CAFs) are known to contribute to cancer progression. We have reported that cell surface expression of hepatocyte growth factor activator inhibitor 1 (HAI-1) is decreased in invasive oral squamous cell carcinoma (OSCC) cells. This study examined if HAI-1-insufficiency contributes to CAF recruitment in OSCC. Serum-free conditioned medium (SFCM) from a human OSCC line (SAS) stimulated the migration of 3 human fibroblast cell lines, NB1RGB, MRC5 and KD. SFCM from HAI-1-knockdown SAS showed an additive effect on the migration of NB1RGB and MRC5, but not KD. SAS SFCM induced protease-activated receptor-2 (PAR-2) expression in NB1RGB and MRC5, but not in KD, and a PAR-2 antagonist blocked the stimulatory effect of HAI-1 knockdown on migration of the PAR-2 expressing cell lines. Moreover, HAI-1-deficient SFCM showed additive stimulatory effects on the migration of wild-type but not PAR-2-deficient mouse fibroblasts. Therefore, the enhanced migration induced by HAI-1-insufficiency was mediated by PAR-2 activation in fibroblasts. This activation resulted from the deregulation of the activity of matriptase, a PAR-2 agonist protease. HAI-1 may thus prevent CAF recruitment to OSCC by controlling matriptase activity. When HAI-1 expression is reduced on OSCC, matriptase may contribute to CAF accumulation by paracrine activation of fibroblast PAR-2. Immunohistochemical analysis of resected OSCC revealed increased PAR2-positive CAFs in 35% (33/95) of the cases studied. The increased PAR-2 positive CAFs tended to correlate with infiltrative histology of the invasion front and shorter disease-free survival of the patients.
已知癌症相关成纤维细胞（CAFs）会促进癌症进展。我们曾报道，侵袭性口腔鳞状细胞癌（OSCC）细胞中肝细胞生长因子激活剂抑制剂1（HAI-1）的细胞表面表达会降低。本研究检测了HAI-1功能不足是否会促进OSCC中CAF的募集。来自人OSCC细胞系（SAS）的无血清条件培养基（SFCM）刺激了3种人成纤维细胞系NB1RGB、MRC5和KD的迁移。来自HAI-1基因敲低的SAS的SFCM对NB1RGB和MRC5的迁移有累加效应，但对KD没有。SAS SFCM在NB1RGB和MRC5中诱导蛋白酶激活受体-2（PAR-2）表达，但在KD中不诱导，并且PAR-2拮抗剂阻断了HAI-1基因敲低对PAR-2表达细胞系迁移的刺激作用。此外，HAI-1缺陷的SFCM对野生型小鼠成纤维细胞的迁移有累加刺激作用，但对PAR-2缺陷的小鼠成纤维细胞没有。因此，HAI-1功能不足诱导的迁移增强是由成纤维细胞中PAR-2激活介导的。这种激活是由PAR-2激动剂蛋白酶matriptase活性失调导致的。因此，HAI-1可能通过控制matriptase活性来阻止CAF募集到OSCC。当OSCC上HAI-1表达降低时，matriptase可能通过旁分泌激活成纤维细胞PAR-2来促进CAF积累。对切除的OSCC进行免疫组织化学分析发现，在所研究的病例中有35%（33/95）的病例中PAR2阳性CAFs增加。PAR-2阳性CAFs增加往往与侵袭前沿的浸润性组织学以及患者较短的无病生存期相关。
