Polarized migration of lymphatic endothelial cells is critically dependent on podoplanin regulation of Cdc42
Polarized migration of lymphatic endothelial cells is critically dependent on podoplanin regulation of Cdc42
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DOI：
10.1152/ajplung.00171.2010
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发表时间：
2011-01-01
影响因子：
4.9
通讯作者：
Ekekezie, Ikechukwu I.
中科院分区：
文献类型：
--
作者：
Navarro, Angels;Perez, Ricardo E.;Ekekezie, Ikechukwu I.
Navarro A, Perez RE, Rezaiekhaligh MH, Mabry SM, Ekekezie II。淋巴内皮细胞的极化迁移严重依赖于podoplanin对Cdc42的调控。[J] .中国生物医学工程学报，2016,31(2):444 - 444。首次发表于2010年10月29日；doi: 10.1152 / ajplung.00171.2010。我们之前已经证明T1 α /podoplanin是人肺微血管淋巴内皮细胞（HMVEC-LLy）形成毛细血管所必需的，并且在淋巴管生成过程开始后不久，podoplanin表达降低的细胞不能正确激活小GTPase RhoA。本研究的目的是确定podoplanin是否调节HMVEC-LLy迁移，以及这种调节是否通过调节小GTPase激活。在抓伤实验分析中，我们发现在HMVEC-LLy中，小干扰RNA （siRNA）缺失podoplanin表达抑制vegf诱导的微管组织中心（MTOC）和高尔基极化，与对照siRNA转染的细胞相比，导致定向迁移显著减少。此外，在这些细胞中观察到皮层肌动蛋白在细胞体纤维网络上的显著重新分布，并且，值得注意的是，如果细胞与Cdc42的显性阴性突变体共转染，它会恢复到控制水平。此外，将Cdc42的显性阴性构建体共转染到podoplanin敲除的HMVEC-LLy中，完全消除了podoplanin缺乏的影响，使MTOC、高尔基极化和细胞迁移恢复到控制水平。重要的是，组成活性Cdc42构建体的表达，如podoplanin敲低，降低了hvec - lly中的RhoA-GTP水平，表明两种gtp酶之间存在串扰。综上所述，结果表明淋巴内皮细胞对VEGF的极化迁移是通过podoplanin调节小GTPase活性，特别是Cdc42的途径介导的。
Navarro A, Perez RE, Rezaiekhaligh MH, Mabry SM, Ekekezie II. Polarized migration of lymphatic endothelial cells is critically dependent on podoplanin regulation of Cdc42. Am J Physiol Lung Cell Mol Physiol 300: L32-L42, 2011. First published October 29, 2010; doi:10.1152/ajplung.00171.2010.-We have shown previously that T1 alpha/podoplanin is required for capillary tube formation by human lung microvascular lymphatic endothelial cells (HMVEC-LLy) and that cells with decreased podoplanin expression fail to properly activate the small GTPase RhoA shortly after the beginning of the lymphangiogenic process. The objective of this study was to determine whether podoplanin regulates HMVEC-LLy migration and whether this regulation is via modulation of small GTPase activation. In analysis of scratch wound assays, we found that small interfering RNA (siRNA) depletion of podoplanin expression in HMVEC-LLy inhibits VEGF-induced microtubule-organizing center (MTOC) and Golgi polarization and causes a dramatic reduction in directional migration compared with control siRNA-transfected cells. In addition, a striking redistribution of cortical actin to fiber networks across the cell body is observed in these cells, and, remarkably, it returns to control levels if the cells are cotransfected with a dominant-negative mutant of Cdc42. Moreover, cotransfection of a dominant-negative construct of Cdc42 into podoplanin knockdown HMVEC-LLy completely abrogated the effect of podoplanin deficiency, rescuing MTOC and Golgi polarization and cell migration to control level. Importantly, expression of constitutively active Cdc42 construct, like podoplanin knockdown, decreased RhoA-GTP level in HMVEC-LLy, demonstrating cross talk between both GTPases. Taken together, the results indicate that polarized migration of lymphatic endothelial cells in response to VEGF is mediated via a pathway of podoplanin regulation of small GTPase activities, in particular Cdc42.