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目的 :研究尿激酶型纤溶酶原激活物 (uPA)及其受体 (uPAR)信号转导对骨巨细胞瘤基质金属蛋白酶 - 2 (MMP - 2 )和金属蛋白酶组织抑制物 - 3(TIMP - 3)的调节。方法 :用免疫组化检测骨巨细胞瘤组织中uPAR、MMP - 2和TIMP - 3的表达。用免疫共沉淀法检测uPA对瘤细胞信号转导通路的p4 4蛋白磷酸化水平。用蛋白印迹法检测用uPA和uPAR抗体处理后瘤细胞MMP - 2和TIMP - 3蛋白表达。结果 :(1)uPAR主要表达在部分单核基质细胞和一些多核巨细胞的胞膜上 ;(2 )MMP - 2主要表达在瘤细胞的胞浆 ,在多核巨细胞 ,其表达有明显的极向性 ;(3)在骨巨细胞瘤组织TIMP - 3表达量低于MMP - 2 ,在多核巨细胞也显示极向性表达 ;(4 )将uPA -ATF加入培养的骨巨细胞瘤细胞后 ,细胞信号通路上的p4 4蛋白磷酸化水平明显增高。用uPAR抗体处理后 ,细胞p4 4蛋白磷酸化水平明显降低。说明uPA -ATF参与细胞信号转导 ,而且受uPAR拮抗剂的影响 ;(5 )uPA -ATF信号通路上调MMP - 2和TIMP - 3的表达 ,而uPAR抗体则下调MMP - 2和TIMP - 3的表达。结论 :本实验首次直接证明uPA -ATF通过信号转导能调节MMP - 2和TIMP - 3的表达 ,而后者则在骨巨细胞瘤局部骨质吸收中起重要作用
AIM: To investigate the effects of urokinase - type plasminogen activator (uPA) and its receptor (uPAR) signaling on the expression of matrix metalloproteinase - 2 (MMP - 2) and tissue inhibitor of metalloproteinase - 3 - 3) adjustment. Methods: The expressions of uPAR, MMP - 2 and TIMP - 3 in giant cell tumor of bone were detected by immunohistochemistry. The level of p4 4 phosphorylation of uPA on tumor cell signal transduction pathway was detected by co-immunoprecipitation. The expressions of MMP - 2 and TIMP - 3 protein in tumor cells were detected by Western blotting with uPA and uPAR antibody. Results: (1) uPAR mainly expressed on the membrane of some mononuclear stromal cells and some multinucleated giant cells. (2) The expression of MMP - 2 was mainly in the cytoplasm of tumor cells. In the multinucleated giant cells, (3) The expression of TIMP - 3 in giant cell tumor of bone is less than that of MMP - 2, and also shows the polar expression in multinucleated giant cells. (4) After adding uPA - ATF into the cultured giant cell tumor , P4 4 protein phosphorylation on the cell signaling pathway was significantly increased. After treatment with uPAR antibody, the level of p4 4 protein phosphorylation was significantly decreased. Indicating that uPA -ATF is involved in cell signal transduction and is influenced by uPAR antagonist; (5) uPA -ATF signal pathway up-regulates the expression of MMP-2 and TIMP-3, while uPAR antibody down-regulates the expression of MMP - 2 and TIMP - 3 expression. CONCLUSIONS: This experiment, for the first time, directly demonstrates that uPA -ATF regulates the expression of MMP-2 and TIMP-3 through signal transduction, whereas the latter plays an important role in the local bone resorption of giant cell tumor of the bone