大肠杆菌血脑屏障侵袭基因 ibeB的稳定转染 能够促进 HeLa 细胞片状伪足的形成
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国家自然科学基金资助项目(30170053, 30170470,30370702).


Expression of ibeB, an Invasion Gene of E.coli Contributing to Crossing of The Blood- brain Barrier, Triggers Lamellipodial Protrusion in HeLa Cells
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This work was supported by a grant from The National Natural Sciences Foundation of China(30170053, 30170470,30370702).

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    摘要:

    为进一步探讨大肠杆菌血脑屏障侵袭基因 ibeB 的生物学特性,将 ibeB 基因克隆至真核表达载体,采用稳定转染的方法,对 ibeB 基因在 HeLa 细胞中的表达所产生的影响进行了研究 . 结果发现:经 ibeB 基因稳定转染的 HeLa 细胞向外周伸展,细胞骨架的组分———微丝发生了重新排布,形成了明显的片状伪足,细胞周边形成较多的膜皱褶结构;细胞对底物的粘附能力和伸展能力显著增强;细胞的粘着斑标志性蛋白 Vinculin 表达增强 . 这些结果为研究片状伪足的形成机制提供了模型 .

    Abstract:

    Several E.coli determinants contributing to invasion of brain microvascular endothelial cells (BMEC) had been previously identified in vitro, including ibeB gene. In order to further characterize the role of the invasion locus ibeB in the absence of other E.coli K1 factors, the ibeB gene was subcloned into eukaryotic expression vector pcDNA3.1/HisC and transfected into HeLa cells. As a result, the HeLa cells expressing ibeB showed dramatic morphological changes comparing to the control, such as the obvious lamellipodial protrusions and membrane ruffles at the periphery of cells due to filamentous actin rearrangement. Cell-substratum adhesion ability and cell spreading ability of ibeB-transfected Hela cells were increased. Furthermore, ibeB could promote the aggregation of vinculin, which play a key role during focal adhesion formation. This first report provides experimental proof to probe the function of ibeB in the process of E.coli K1 invasion and the mechanism of lamellipodial protrusion.

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赵伟东,陈誉华.大肠杆菌血脑屏障侵袭基因 ibeB的稳定转染 能够促进 HeLa 细胞片状伪足的形成[J].生物化学与生物物理进展,2005,32(2):116-121

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