Phospholipase D1 regulates cell migration in a lipase activity-independent manner
Phospholipase D1 regulates cell migration in a lipase activity-independent manner
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DOI：
10.1074/jbc.m509844200
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发表时间：
2006-06-09
影响因子：
4.8
通讯作者：
Ryu, Sung Ho
中科院分区：
文献类型：
--
作者：
Kim, Jung Hwan;Kim, Hyun-Wook;Ryu, Sung Ho
细胞迁移是一个复杂的生物过程，需要动态的细胞骨架重塑。磷脂酶D （PLD）产生磷脂酸，磷脂酸是脂质的第二信使。尽管PLD活性已被提出在细胞骨架重排中发挥作用，但PLD参与重排过程的方式仍然不清楚。在这项研究中，通过在HeLa细胞中使用小干扰RNA沉默内源性PLD1同工酶，我们证明内源性PLD1对于肌动蛋白细胞骨架的正常组织是必需的，更重要的是，对于细胞运动是必需的。在HeLa细胞中，PLD1的沉默导致细胞形态的剧烈变化，包括应力纤维的积累，以及细胞伸长和变平，这似乎是由局灶粘连数量增加引起的，最终导致细胞-基质相互作用增强。因此，血清诱导的细胞迁移被pld1沉默严重抑制。此外，通过向敲除细胞中添加野生型和脂肪酶无活性PLD1，可以恢复PLD1沉默引起的细胞基质相互作用增强和细胞迁移迟缓。综上所述，我们的研究结果强烈表明，内源性PLD1是基于肌动蛋白的细胞骨架组织的关键因素，涉及细胞粘附和迁移。PLD1的这些作用似乎以不依赖于脂肪酶活性的方式起作用。我们还讨论了PLD1对Src家族激酶的调节，这与Pyk2和细胞迁移的调节有关。
Cell migration, a complex biological process, requires dynamic cytoskeletal remodeling. Phospholipase D (PLD) generates phosphatidic acid, a lipid second messenger. Although PLD activity has been proposed to play a role in cytoskeletal rearrangement, the manner in which PLD participates in the rearrangement process remains obscure. In this study, by silencing endogenous PLD isozymes using small interfering RNA in HeLa cells, we demonstrate that endogenous PLD1 is required for the normal organization of the actin cytoskeleton, and, more importantly, for cell motility. PLD1 silencing in HeLa cells resulted in dramatic changes in cellular morphology, including the accumulation of stress fibers, as well as cell elongation and flattening, which appeared to be caused by an increased number of focal adhesions, which ultimately culminated in enhanced cell-substratum interactions. Accordingly, serum-induced cell migration was profoundly inhibited by PLD1-silencing. Moreover, the augmented cell substratum interaction and retarded cell migration induced by PLD1-silencing could be restored by the adding back not only of wild type, but also of lipase-inactive PLD1 into knockdown cells. Taken together, our results strongly suggest that endogenous PLD1 is a critical factor in the organization of the actin-based cytoskeleton, with regard to cell adhesion and migration. These effects of PLD1 appear to operate in a lipase activity-independent manner. We also discuss the regulation of Src family kinases by PLD1, as related to the modulation of Pyk2 and cell migration.