【摘要】 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。更多还原

【Abstract】 Objective To investigate the effects of β1-integrin,fibronectin(FN) and laminin(LN) on microfilament and glial fibrillary acidic protein( GFAP) of U251 malignant glioma cells. Methods The amount and distributions of microfilaments and GFAP of U251MG cells were observed. The effects of β1-integrin,FN and LN on microfilament skeleton and GFAP were analysed with immunoflrorescen cytochemistry and confocal laser scanning microscope. The effects of them on migration capacity of U251MG cells were detected by migration assay in vitro. Results The microfilament network form strong,dense and orderly arranged structure in groups added FN and LN. The anti-β1integrin antibodies led to disruption of microfilament network as well as presence of F-actin aggregates. The anti-β1integrin antibodies reduced the GFAP expression. FN and LN showed no effect on the expression and structure of GFAP. Co-expression of microfilament and GFAP in U251MG cells was F-actin. The cell migration of U251MG cells on dishes coated with FN and LN was inhibited by anti-β1integrin antibodies( P < 0. 05). Conclusion The interaction between β1-integrin,FN and LN may change the number and the structure of microfilament skeleton and GFAP of U251MG cell to promote its migration ability. The structure of GFAP in U251MG cells is related to the F-actin.更多还原