Myosin light chain kinase mediates transcellular intravasation of breast cancer cells through the underlying endothelial cells: a three-dimensional FRET study
Myosin light chain kinase mediates transcellular intravasation of breast cancer cells through the underlying endothelial cells: a three-dimensional FRET study
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DOI：
10.1242/jcs.053793
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发表时间：
2010-02-01
影响因子：
4
通讯作者：
Chew, Teng-Leong
中科院分区：
文献类型：
--
作者：
Khuon, Satya;Liang, Luke;Chew, Teng-Leong
侵袭性乳腺癌细胞和底层内皮细胞之间的瞬时和局部信号传导事件的特征仍然很差。我们报告了一种新的方法，将血管工程与三维时间推移荧光共振能量转移（FRET）成像相结合，以剖析内皮肌球蛋白轻链激酶（MLCK）在肿瘤内渗过程中是如何被调节的。我们表明，肿瘤跨内皮迁移发生通过细胞旁（即通过细胞-细胞连接）和跨细胞（即通过单个内皮细胞）的路线。内皮MLCK在侵袭部位被激活，导致肌球蛋白-II调节轻链（RLC）的区域二磷酸化和肌球蛋白收缩。阻断内皮RLC二磷酸化减弱肿瘤的跨细胞侵袭，但不减弱细胞旁侵袭。我们的研究结果暗示了内皮肌球蛋白-II功能在肿瘤内渗中的重要作用。
The transient and localized signaling events between invasive breast cancer cells and the underlying endothelial cells have remained poorly characterized. We report a novel approach integrating vascular engineering with three- dimensional time-lapse fluorescence resonance energy transfer (FRET) imaging to dissect how endothelial myosin light chain kinase (MLCK) is modulated during tumor intravasation. We show that tumor transendothelial migration occurs via both paracellular (i.e. through cell-cell junctions) and transcellular (i.e. through individual endothelial cells) routes. Endothelial MLCK is activated at the invasion site, leading to regional diphosphorylation of myosin-II regulatory light chain (RLC) and myosin contraction. Blocking endothelial RLC diphosphorylation blunts tumor transcellular, but not paracellular, invasion. Our results implicate an important role for endothelial myosin-II function in tumor intravasation.