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目的研究整合素β1和细胞外基质[主要选取层黏连蛋白(laminin,LN)和纤黏连蛋白(fibronectin,FN)多形性胶质母细胞瘤U251细胞(U251MG)]对细胞微丝和中间丝(胶质纤维酸性蛋白glial fibrillary acidic protein,GFAP)骨架的影响。方法体外整合素β1抗体和细胞外基质处理U251MG细胞系,通过免疫荧光细胞化学染色及激光共聚焦显微镜,观察并分析细胞微丝和GFAP细胞骨架在细胞中定位的变化;通过运动迁移实验分析其对细胞运动能力的影响。结果 FN、LN包被处理的U251MG细胞内可见粗壮而密集、清晰的微丝结构。抗整合素β1抗体处理的细胞内,微丝分布杂乱而模糊,并可见大量絮团状的F-actin小体。抗整合素β1抗体处理的细胞内GFAP染色荧光信号减弱,FN和LN对GFAP表达及结构无影响。微丝和GFAP双标均阳性重合处位于F-actin小体处。抗整合素β1抗体减弱U251MG细胞在FN和LN的运动、迁移能力(P<0.05)。结论在U251MG细胞中,通过整合素β1和FN、LN相互作用,可改变细胞微丝的分布,并以此影响胶质瘤细胞的运动和迁移。GFAP在U251MG细胞内的分布同F-actin小体有关。
Objective To investigate the effects of integrin β1 and extracellular matrix (U251MG), a major selective laminin (LN) and fibronectin (FN) glioblastoma multiforme, Effect of intermediate filament (glial fibrillary acidic protein, GFAP) scaffolds. Methods The U251MG cell line was treated with integrin β1 antibody and extracellular matrix in vitro. The immunofluorescence staining and confocal laser scanning microscopy were used to observe the changes of the localization of the actin cytoskeleton and GFAP cytoskeleton in the cells. Impact on cell motility. Results FN, LN coated U251MG cells can be seen thick, dense, clear microfilament structure. Anti-integrin β1 antibody-treated cells, microfilament distribution messy and fuzzy, and visible a large number of flocculent F-actin bodies. Anti-integrinβ1 antibody treated intracellular GFAP staining fluorescence signal decreased, FN and LN had no effect on GFAP expression and structure. Microfilament and GFAP double-labeled positive coincidence at the F-actin body. Anti-integrin β1 antibody attenuated the motility and migration ability of U251MG cells in FN and LN (P <0.05). Conclusion U251MG cells can change the distribution of microfilament through the interaction between integrin β1 and FN and LN, and thus affect the movement and migration of glioma cells. The distribution of GFAP in U251MG cells was related to F-actin bodies.