Protein C inhibitor regulates both cathepsin L activity and cell-mediated tumor cell migration
Protein C inhibitor regulates both cathepsin L activity and cell-mediated tumor cell migration
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DOI：
10.1016/j.bbagen.2010.03.003
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发表时间：
2010-06-01
影响因子：
3
通讯作者：
Church, Frank C.
中科院分区：
文献类型：
--
作者：
Fortenberry, Yolanda M.;Brandal, Stephanie;Church, Frank C.
背景资料：蛋白C抑制剂（PCI）是一种血浆丝氨酸蛋白酶抑制剂（serpin），可调节凝血过程中的几种丝氨酸蛋白酶，包括凝血酶和活化蛋白C。然而，PCI的生理作用仍在研究中。半胱氨酸蛋白酶，组织蛋白酶L在许多生理过程中起作用，包括心血管疾病，血管重塑和cancer.Methods和结果：我们发现PCI抑制组织蛋白酶L，抑制率（k（2））为3.0 × 10（5）M（-1）s（-1）。然而，PCI P1突变体（R354 A）以与野生型PCI相似的速率抑制组织蛋白酶L，突变P2残基导致抑制速率略微降低。然后，我们评估了PCI和组织蛋白酶L对人乳腺癌（MDA-MB-231）细胞迁移的影响。组织蛋白酶L在MDA-MB-231细胞的细胞裂解物和条件培养基中均表达。外源性给予wtPCI和PCI P1可抑制MDA-MB-231细胞的创伤诱导迁移和跨孔迁移，但PCI P14突变体不抑制。此外，与不表达的MDA-MB-231细胞或表达PCI P14突变体的MDA-MB-231细胞相比，表达wtPCI的MDA-MB-231细胞的迁移显著降低。下调组织蛋白酶L的特异性组织蛋白酶L抑制剂或siRNA技术也导致在MDA-MB-231 cells.Conclusions的迁移减少：总体而言，我们的数据表明，PCI调节肿瘤细胞迁移部分通过抑制组织蛋白酶L。一般意义：因此，抑制组织蛋白酶L的丝氨酸蛋白酶抑制剂像PCI可能是一个新的途径，调节止血，心血管和转移性疾病。(C)2010 Elsevier B. V.保留所有权利。
Background: Protein C inhibitor (PCI) is a plasma serine protease inhibitor (serpin) that regulates several serine proteases in coagulation including thrombin and activated protein C. However, the physiological role of PCI remains under investigation. The cysteine protease, cathepsin L has a role in many physiological processes including cardiovascular diseases, blood vessel remodeling, and cancer.Methods and results: We found that PCI inhibits cathepsin L with an inhibition rate (k(2)) of 3.0 x 10(5) M(-1) s(-1). Whereas, the PCI P1 mutant (R354A) inhibits cathepsin L at rates similar to wild-type PCI, mutating the P2 residue results in a slight decrease in the rate of inhibition. We then assessed the effect of PCI and cathepsin L on the migration of human breast cancer (MDA-MB-231) cells. Cathepsin L was expressed in both the cell lysates and conditioned media of MDA-MB-231 cells. Wound-induced and transwell migration of MDA-MB-231 cells was inhibited by exogenously administered wtPCI and PCI P1 but not PCI P14 mutant. In addition, migration of MDA-MB-231 cells expressing wtPCI was significantly decreased compared to non-expressing MDA-MB-231 cells or MDA-MB-231 cells expressing the PCI P14 mutant. Downregulation of cathepsin L by either a specific cathepsin L inhibitor or siRNA technology also resulted in a decrease in the migration of MDA-MB-231 cells.Conclusions: Overall, our data show that PCI regulates tumor cell migration partly by inhibiting cathepsin L.General significance: Consequently, inhibiting cathepsin L by serpins like PCI may be a new pathway of regulating hemostasis, cardiovascular and metastatic diseases. (C) 2010 Elsevier B.V. All rights reserved.