Helicobacter pylori VacA cytotoxin interacts with fibronectin and alters HeLa cell adhesion and cytoskeletal organization in vitro

Ewa E. Hennig , Michał M. Godlewski , Eugeniusz Butruk , Jerzy Ostrowski
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引用次数: 33

Abstract

Helicobacter pylori vacuolating cytotoxin VacA causes multiple effects on epithelial cell function and morphology, but the effects of VacA on signal transduction pathways and the cytoskeleton have not been investigated in detail. In this study, we analyzed the effects of native VacA on HeLa and AGS cell adhesion to fibronectin and laminin under serum-free conditions. Confocal microscopic examination revealed increased number of cells with rounded morphology and inhibition of actin fiber formation, in the presence of VacA. VacA binds to fibronectin in vitro in a dose-dependent manner. This interaction was partly inhibited by a peptide containing an arginine-glycine-aspartic acid motif. The adhesion of HeLa cells to fibronectin, but not to laminin, was decreased in the presence of VacA. Thus, VacA may interact with fibronectin and influence integrin receptor-induced cell signaling and cytoskeleton-dependent cell functions.

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幽门螺杆菌VacA细胞毒素与纤维连接蛋白相互作用并改变HeLa细胞粘附和细胞骨架组织
幽门螺杆菌空泡细胞毒素VacA对上皮细胞的功能和形态有多种影响,但VacA对信号转导通路和细胞骨架的影响尚未得到详细研究。在本研究中,我们分析了在无血清条件下,天然VacA对HeLa和AGS细胞对纤维连接蛋白和层粘连蛋白粘附的影响。共聚焦显微镜检查显示,在VacA存在的情况下,圆形形态的细胞数量增加,肌动蛋白纤维形成受到抑制。VacA在体外以剂量依赖的方式与纤维连接蛋白结合。这种相互作用被含有精氨酸-甘氨酸-天冬氨酸基序的肽部分抑制。在VacA存在的情况下,HeLa细胞对纤维连接蛋白的粘附能力下降,而对层粘连蛋白的粘附能力没有下降。因此,VacA可能与纤维连接蛋白相互作用,影响整合素受体诱导的细胞信号传导和细胞骨架依赖的细胞功能。
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