ECM dependent and integrin mediated tumor cell migration of human glioma and melanoma cell lines under serum-free conditions.
ECM dependent and integrin mediated tumor cell migration of human glioma and melanoma cell lines under serum-free conditions.
复制标题
在无血清条件下，ECM 依赖性和整合素介导的人神经胶质瘤和黑色素瘤细胞系的肿瘤细胞迁移。
作者：
R. Goldbrunner;H. K. Haugland;C. Klein;S. Kerkau;K. Roosen;J. Tonn
IV型胶原、层粘连蛋白和纤维连接蛋白是脑细胞外基质（ECM）的组成成分，在胶质瘤细胞侵袭中起关键作用。本研究的目的是评估两种不同侵袭性肿瘤在无基质条件下整合素依赖的细胞-基质相互作用。两种人胶质瘤（GaMG, U373）和黑色素瘤（MV3， BLM）细胞系在无血清培养基中生长。免疫荧光显微镜观察IV型胶原蛋白、层粘连蛋白和纤维连接蛋白。定量分析了这些ECM成分对单层细胞的粘附和向多细胞球体的迁移。已知作为层粘连蛋白受体的整合素链在额外的迁移试验中被特异性抗体阻断。在无血清条件下，所有细胞系均表达所有ECM成分。在胶质瘤和黑色素瘤细胞系中，所有ECM成分都增加了肿瘤细胞的粘附和迁移，其中层粘连蛋白是最强的迁移促进剂。然而，胶质瘤的迁移是剂量依赖性的，而黑色素瘤显示10微克/毫升层粘连蛋白的最佳剂量。抗- 3整合素抗体在所有细胞系中显著减少层粘连蛋白的迁移，抗- 1抗体在除U373外的所有细胞系中显著减少层粘连蛋白的迁移。抗- α 2在BLM中表现出较强的作用，抗- α 6在胶质瘤中比在黑色素瘤中表现出更强的抑制作用。整合素在层粘连蛋白上参与肿瘤细胞的功能运动。层粘连蛋白相关整合素链的阻断以不同的方式显著降低细胞系之间的细胞运动性。此外，不同细胞系利用不同的整合素作为层粘连蛋白受体。
Collagen IV, laminin and fibronectin are constituents of the cerebral extracellular matrix (ECM), which is critical in glioma cell invasion. The aim of the present study was to evaluate the integrin dependent cell-matrix interactions of two tumors with different invasive properties under matrixfree conditions. Two human glioma (GaMG, U373) and melanoma (MV3, BLM) cell lines were grown in serum free medium. Immunofluorescence microscopy of collagen IV, laminin, and fibronectin was performed. The adhesion of monolayer cells and their migration out of multicellular spheroids was quantified for these ECM components. Integrin chains known to act as laminin receptors were blocked by specific antibodies in additional migration assays. All cell lines expressed all the ECM components under serum free conditions. Tumor cell adhesion and migration in both glioma and melanoma cell lines was increased by all the ECM components, laminin being the strongest promotor of migration. However, migration was dose dependent in gliomas, whereas melanomas revealed a dose optimum of 10 micrograms/ml laminin. Antibodies against alpha 3 integrins significantly reduced migration on laminin in all cell lines, anti-beta 1 in all cell lines except U373. Anti-alpha 2 in BLM showed a strong effect, anti-alpha 6 was a stronger inhibitor in glioma than in melanoma cells. Integrins are functionally involved in tumor cell locomotion on laminin. The blocking of laminin related integrin chains markedly reduces cell motility in a varying manner between the cell lines. Moreover, different cell lines utilize different integrins as the laminin receptor.