Itatsu Keita, Sasaki Motoko, Yamaguchi Junpei, Ohira Shusaku, Ishikawa Akira, Ikeda Hiroko, Sato Yasunori, Harada Kenichi, Zen Yoh, Sato Hiroshi, Ohta Tetsuo, Nagino Masato, Nimura Yuji, Nakanuma Yasuni
Department of Human Pathology, Kanazawa University Graduate School of Medicine, Kanazawa 920-8640, Japan.
Am J Pathol. 2009 Mar;174(3):829-41. doi: 10.2353/ajpath.2009.080012. Epub 2009 Feb 13.
Matrix metalloproteinase-9 (MMP-9) is an important enzyme in tumor invasion and metastasis in malignant tumors, including cholangiocarcinoma (CC). Tumor necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine, was recently reported to induce the up-regulation of MMP-9 in cultured CC cells. We examined whether cyclooxygenase-2 (COX-2) and prostaglandin-E2 (PGE2), another endogenous tumor promoter, are involved in the up-regulation of MMP-9 in CC using CC tissue specimens and a CC cell line, HuCCT-1. MMP-9 and COX-2 were immunohistochemically expressed in 58% and 89% of 110 CC cases, respectively; the expression of MMP-9 and COX-2 was correlated (r = 0.32, P = 0.00072). Using zymography, latent MMP-9 was detectable in all cases and active MMP-9 was detected in 24% of cases of the CC specimens. The TNF-alpha/TNF-receptor 1 (TNF-R1) interaction induced MMP-9 production and activation, as well as COX-2 overexpression and PGE2 production, and increased the migration of CC cells. MMP-9 up-regulation was inhibited by COX inhibitors, antagonists of EP2/4 (receptors of PGE2), and COX-1 and COX-2 siRNAs. Inhibitors of both MMP-9 and MMP-9 siRNA treatment abrogated the increase in the migration of CC cells induced by TNF-alpha. In conclusion, we propose a novel signaling pathway of MMP-9 up-regulation in CC cells such that TNF-alpha induces the activation of COX-2 and PGE2 via TNF-R1 followed by the up-regulation of MMP-9 via the PGE2 (EP2/4) receptor.
基质金属蛋白酶-9（MMP-9）是包括胆管癌（CC）在内的恶性肿瘤中肿瘤侵袭和转移的一种重要酶。肿瘤坏死因子-α（TNF-α）是一种促炎细胞因子，最近有报道称其可诱导培养的CC细胞中MMP-9上调。我们使用CC组织标本和CC细胞系HuCCT-1，研究了另一种内源性肿瘤促进因子环氧合酶-2（COX-2）和前列腺素-E2（PGE2）是否参与CC中MMP-9的上调。在110例CC病例中，MMP-9和COX-2的免疫组化表达分别为58%和89%；MMP-9和COX-2的表达呈正相关（r = 0.32，P = 0.00072）。通过酶谱分析，在所有病例中均可检测到潜伏性MMP-9，在CC标本的24%病例中检测到活性MMP-9。TNF-α/TNF受体1（TNF-R1）相互作用诱导MMP-9的产生和激活，以及COX-2的过表达和PGE2的产生，并增加CC细胞的迁移。MMP-9的上调受到COX抑制剂、EP2/4（PGE2受体）拮抗剂以及COX-1和COX-2 siRNA的抑制。MMP-9抑制剂和MMP-9 siRNA处理均消除了TNF-α诱导的CC细胞迁移增加。总之，我们提出了一种CC细胞中MMP-9上调新的信号通路，即TNF-α通过TNF-R1诱导COX-2和PGE2的激活，随后通过PGE2（EP2/4）受体上调MMP-9。
