Department of Biophysics, Medical Centre for Postgraduate Education, Warsaw, Poland.
Cell Biol Int. 2012 Oct 1;36(10):883-92. doi: 10.1042/CBI20110522.
Interaction of cell integrins with the ECM (extracellular matrix) proteins is commonly assumed to be associated with cell dissemination and tumour metastases. Since these processes depend on the mechanism of cell-protein interaction, we have attempted to show the contribution of α5β1 and αvβ3 integrins of the prostate cancer PC-3 cells in in vitro interaction with FN (fibronectin) adsorbed on defined polystyrene surfaces. Cell adhesion, spreading and cytoskeleton organization were studied using antibodies against integrins or a GRGDSP (Gly-Arg-Gly-Asp-Ser-Pro) peptide. The results show that blocking the α5β1 integrin causes: (i) a decrease in the number of the adherent cells in the early phase of adhesion and (ii) a decrease in the dynamics of cell spreading and cell shape changes, and weaker reorganization of cytoskeletal proteins than in the control cells. Conversely, the blocking of the αvβ3 integrin: (i) causes no observable effect on the number of the adhered cells; however, (ii) causes an increase in the dynamics of cell spreading and cell shape changes, and stronger reorganization of cytoskeletal proteins than in the control cells. Interestingly, the blocking of integrins with a GRGDSP peptide strongly decreases the number of the adhered cells, and a complete inhibition of cell spreading. Our results strongly suggest that the α5β1 integrin plays the main role in the adhesion and spreading of PC-3 cells interacting with FN, whereas the αvβ3 integrin seems to regulate other receptors in the spreading process. Moreover, integrin-FN interaction through the RGD sequence evidently curbed the cell adhesion and spreading.
细胞整合素与细胞外基质（ECM）蛋白的相互作用通常被认为与细胞扩散和肿瘤转移有关。由于这些过程依赖于细胞-蛋白相互作用的机制，我们试图展示前列腺癌细胞 PC-3 中α5β1 和αvβ3 整合素在体外与吸附在特定聚苯乙烯表面上的 FN（纤连蛋白）相互作用中的作用。使用针对整合素或 GRGDSP（甘氨酰-精氨酰-甘氨酰-天冬氨酸-丝氨酸-脯氨酸）肽的抗体研究了细胞黏附、铺展和细胞骨架组织。结果表明，阻断α5β1 整合素会导致：（i）在黏附的早期阶段，附着细胞的数量减少；（ii）细胞铺展和细胞形状变化的动力学减少，细胞骨架蛋白的重组比对照细胞弱。相反，阻断αvβ3 整合素：（i）对附着细胞的数量没有可观察到的影响；但是，（ii）导致细胞铺展和细胞形状变化的动力学增加，细胞骨架蛋白的重组比对照细胞强。有趣的是，用 GRGDSP 肽阻断整合素强烈降低附着细胞的数量，并完全抑制细胞铺展。我们的结果强烈表明，α5β1 整合素在与 FN 相互作用的 PC-3 细胞的黏附和铺展中起主要作用，而αvβ3 整合素似乎在铺展过程中调节其他受体。此外，通过 RGD 序列的整合素-FN 相互作用显然抑制了细胞黏附和铺展。
