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Huang, Yi-Wei; Chang, Shu-Jing; I-Chen Harn, Hans; Huang, Hui-Ting; Lin, Hsi-Hui; Shen, Meng-Ru; Tang, Ming-Jer; Chiu, Wen-Tai
Journal of cellular physiology, 09/2015, Volume: 230, Issue: 9Journal Article
Ca2+‐mediated formation of cell polarity is essential for directional migration which plays an important role in physiological and pathological processes in organisms. To examine the critical role of store‐operated Ca2+ entry, which is the major form of extracellular Ca2+ influx in non‐excitable cells, in the formation of cell polarity, we employed human bone osteosarcoma U2OS cells, which exhibit distinct morphological polarity during directional migration. Our analyses showed that Ca2+ was concentrated at the rear end of cells and that extracellular Ca2+ influx was important for cell polarization. Inhibition of store‐operated Ca2+ entry using specific inhibitors disrupted the formation of cell polarity in a dose‐dependent manner. Moreover, the channelosomal components caveolin‐1, TRPC1, and Orai1 were concentrated at the rear end of polarized cells. Knockdown of TRPC1 or a TRPC inhibitor, but not knockdown of Orai1, reduced cell polarization. Furthermore, disruption of lipid rafts or overexpression of caveolin‐1 contributed to the downregulation of cell polarity. On the other hand, we also found that cell polarity, store‐operated Ca2+ entry activity, and cell stiffness were markedly decreased by low substrate rigidity, which may be caused by the disorganization of actin filaments and microtubules that occurs while regulating the activity of the mechanosensitive TRPC1 channel. J. Cell. Physiol. 230: 2086–2097, 2015. © 2015 Wiley Periodicals, Inc.
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