尿激酶型纤溶酶原激活物系统对骨巨细胞瘤细胞基质金属蛋白酶-2和组织基质金属蛋白酶抑制物-3表达的影响  

Regulation of MMP-2 and TIMP-3 expression by uPA signal transduction system in human bone giant cell tumor

作　　者：徐若冰[1] 文剑明[2] 张萌[2] 吕长海[3] 肖刚[2] 张文敏[2] 梁惠珍[2] 

机构地区：[1]广州医学院病理学教研室,广东广州510182 [2]中山大学中山医学院病理学教研室,广东广州510080 [3]中山大学附属口腔医院儿童牙科,广东广州510060

出　　处：《中国病理生理杂志》2004年第11期1982-1988,共7页Chinese Journal of Pathophysiology

基　　金：教育部高等学校博士学科点专项科研基金资助项目 (No.2 0 0 0 4 6 )

摘　　要：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 。AIM: To study effects of urokinase-type plasminogen activator (uPA) signal transduction on expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) in giant cell tumor of bone (GCT). METHODS: Expression of uPAR, MMP-2 and TIMP-3 in GCT tissue was detected by immunohistochemistry. Phosphorylation level of mitogen-activated protein kinase (p44) in uPA/uPAR signal pathway in cultured GCT cells was detected by immunoprecipitation. The expression of MMP-2 and TIMP-3 in cultured cells after treatment with uPA-ATF or anti-uPAR antibody was also detected by Western blotting. RESULTS: 1) Urokinase-type plasminogen activator receptor (uPAR) was positive on the cell membrane and in cytoplasm of some mononuclear stromal cells (MSCs) and multinucleated giant cells (MGCs); 2) MMP-2 was positive in the cytoplasm and on the cell membrane of almost all of MSCs and some of MGCs. The polar distribution of MMP-2 in the cytoplasm of MGCs was especially obvious; 3) The expression of TIMP-3 of some MSCs and MGCs in GCT was much lower than MMP-2. The positive signal also showed a prominent polarity; 4) After treatment with uPA-ATF, the phosphorylation level of p44 in GCT cultured cells was much higher than the control. Addition of anti-uPAR antibody in the cells remarkably down-regulated the phosphorylation level of p44 as compared with the control group, suggesting that uPA-ATF participates cell signal transduction and this reaction can be inhibited by anti-uPAR antibody; 5) uPA-ATF cell signal pathway up-regulated expression of MMP-2 and TIMP-3, while anti-uPAR antibody down-regulated the expression of MMP-2 and TIMP-3. CONCLUSION: These results demonstrate for the first time that uPA-ATF directly regulates the expression of MMP-2 and TIMP-3 by signal transduction pathway, and the over-expression of MMP-2 and TIMP-3 may play an important role in local osteolysis of GCT. [

关 键 词：巨细胞瘤 骨 尿纤溶酶原激活物 信号转导 

分 类 号：R363[医药卫生—病理学]