MMP-2对人源黑色素瘤A375细胞粘附和迁移影响的研究

发布时间：2018-09-10 11:54

【摘要】： 恶性黑色素瘤因具有高度迁移性和致死性而被人们熟知。明胶酶,即基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9),在黑色素瘤迁移过程中参与基底膜降解和细胞外基质重构,与肿瘤恶性化程度密切相关。明胶酶不仅参与胞外基质与基底膜重构,在血管发生、侵润和迁移等肿瘤发生过程中发挥了重要作用。随着对明胶酶研究的深入,人们发现明胶酶不仅由肿瘤细胞分泌并且可以由基质细胞分泌,不仅能与细胞外基质蛋白相互作用,而且能与包括整合素在内的多种非基质蛋白相互作用。在黑色素瘤中,整合素αvβ3(integrin)是一类重要的异二聚体跨膜糖蛋白,能够与含有RGD片段的配体结合,通过这种结合αvβ3可以介导细胞外配体与胞内骨架蛋白和各种信号途径间的信号传递。然而,整合素αvβ3与MMP-2间的识别是通过MMP-2羧基端的PEX区而非RGD区,对于MMP-2是否能通过与αvβ3的这种结合而发挥一定的信号传递作用目前还没有报道。在本研究中,我们首先利用明胶酶谱法对A375细胞中明胶酶的表达进行了检测,证实在人源黑色素瘤A375细胞中MMP-2和MMP-9均有所表达。通过使用MMP广谱抑制剂对A375细胞铺展和迁移进行观察发现,MMP-2对A375细胞的影响比较显著。通过对A375细胞铺展过程中MMP-2和F-actin进行免疫荧光标记,发现MMP-2与肌动蛋白细胞骨架的变化有一定相关性。同时,免疫荧光标记观察发现MMP-2和整合素αvβ3在A375细胞悬浮、铺展和迁移三种状态下均有共定位现象。通过静止粘附实验我们证实,MMP-2能够影响整合素αvβ3与其配体的结合能力。这些结果表明,MMP-2在A375中可能通过整合素αvβ3而发挥某些信号作用从而影响肿瘤细胞的发展。
[Abstract]:Malignant melanoma is well known for its high mobility and mortality. Gelatinase, namely matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9), is involved in basement membrane degradation and extracellular matrix remodeling during melanoma migration, which is closely related to the malignancy of the tumor. Gelatinase not only participates in the remodeling of extracellular matrix and basement membrane, but also plays an important role in angiogenesis, invasion and migration. With the further study of gelatinase, it has been found that gelatinase is not only secreted by tumor cells but also by stromal cells, and can interact with extracellular matrix proteins. And can interact with a variety of non-matrix proteins, including integrins. In melanoma, integrin 伪 v 尾 3 (integrin) is an important heterodimer transmembrane glycoprotein that binds to ligand containing RGD fragments. This binding 伪 v 尾 3 mediates signal transduction between extracellular ligands, intracellular cytoskeleton proteins and various signal pathways. However, the recognition of integrin 伪 v 尾 3 and MMP-2 is through the PEX region of the carboxyl terminal of MMP-2 rather than the RGD region. It has not been reported whether MMP-2 can play a certain role in signal transduction through the combination of 伪 v 尾 3 with 伪 v 尾 3. In this study, we first detected the expression of gelatinase in A375 cells by gelatinase spectroscopy, and confirmed that both MMP-2 and MMP-9 were expressed in human melanoma A375 cells. The proliferation and migration of A375 cells were observed by using MMP broad-spectrum inhibitor. It was found that MMP-2 had a significant effect on A375 cells. By immunofluorescence labeling of MMP-2 and F-actin during the spreading of A375 cells, it was found that MMP-2 was related to the change of actin cytoskeleton. At the same time, immunofluorescence staining showed that MMP-2 and integrin 伪 v 尾 3 were co-located in A375 cell suspension, spreading and migration. Through the static adhesion experiment, we confirmed that MMP-2 can affect the binding ability of integrin 伪 v 尾 3 to its ligand. These results suggest that MMP-2 may play some signaling role in A375 by integrin 伪 v 尾 3, which may affect the development of tumor cells.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.5

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