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Roles of Serine/Threonine Phosphatases in Low-Dose Endothelial Monocyte-Activating Polypeptide-II-Induced Opening of Blood–Tumor Barrier

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Abstract

Previous studies have demonstrated that low-dose endothelial monocyte-activating polypeptide-II (EMAP-II) induces blood–tumor barrier (BTB) opening via RhoA/Rho kinase/PKC-α/β signaling pathway. In a recent study, we revealed that low-dose EMAP-II induced significant increases in expression levels of serine/threonine (Ser/Thr) phosphatase (PP)1 and 2A in rat brain microvascular endothelial cells (RBMECs) of BTB model. In addition, PKC-ζ/PP2A signaling pathway is involved in EMAP-II-induced BTB hyperpermeability. The present study further investigated the exact roles of PPs in this process. In an in vitro BTB model, low-dose EMAP-II (0.05 nM) induced a significant increase in PP1 activity in RBMECs. There was an interaction between PKC-α/β and PP1 in RBMECs. Inhibition of PKC-α/β activity with GÖ6976 completely blocked EMAP-II-induced activation of PP1. Conversely, inhibition of PP1 activity with tautomycin had no effect on EMAP-II-induced PKC-α/β activation. Like GÖ6976, tautomycin significantly prevented EMAP-II-induced BTB hyperpermeability and MLC phosphorylation in RBMECs. Also, in this study, EMAP-II induced a marked redistribution of occludin and a significant dephosphorylation of occludin on Ser/Thr residues in RBMECs. Similar with GÖ6976 pretreatment, tautomycin pretreatment dramatically diminished EMAP-II-induced redistribution of occludin. Furthermore, pretreatment with tautomycin significantly inhibited EMAP-II-induced dephosphorylation of occludin on Ser residues. However, pretreatment with okadaic acid (an inhibitor of PP2A) significantly prevented changes in Ser-phosphorylated occludin induced by EMAP-II treatment. Collectively, this study demonstrates that low-dose EMAP-II increases BTB permeability via a RhoA/Rho kinase/PKC-α/β/PP1 signaling pathway and that PP1/PP2A-mediated Ser/Thr dephosphorylation of occludin plays an important role in EMAP-II-induced BTB hyperpermeability.

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Acknowledgments

This work is supported by grants from the Natural Science Foundation of China (Nos. 81372484, 81372682, 81172197, 81171131, 81272564, and 81272795), the Natural Science Foundation of Liaoning Province in China (No. 201102300), Liaoning Science and Technology Plan Projects (No. 2011225020), Shenyang Science and Technology Plan Projects (Nos. F11-264-1-15, F12-277-1-05, F13-318-1-16, F13-318-1-19 and F13-220-9-15), and Outstanding Scientific Fund of Shengjing Hospital (No. 201304).

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Authors and Affiliations

  1. Department of Neurosurgery, Shengjing Hospital, China Medical University, Shenyang, Liaoning Province, 110004, People’s Republic of China

    Zhen Li, Yun-hui Liu, Jing Liu & Yi-long Yao

  2. The 96th Class, 7-Year Program, China Medical University, Shenyang, Liaoning Province, 110001, People’s Republic of China

    Xiao-bai Liu

  3. Department of Neurobiology, College of Basic Medicine, China Medical University, Shenyang, Liaoning Province, 110001, People’s Republic of China

    Yi-xue Xue, Ping Wang, Li-bo Liu & Jun Ma

  4. Institute of Pathology and Pathophysiology, China Medical University, Shenyang, Liaoning Province, 110001, People’s Republic of China

    Yi-xue Xue, Ping Wang, Li-bo Liu & Jun Ma

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  1. Zhen Li
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  2. Xiao-bai Liu
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Correspondence to Yun-hui Liu.

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Li, Z., Liu, Xb., Liu, Yh. et al. Roles of Serine/Threonine Phosphatases in Low-Dose Endothelial Monocyte-Activating Polypeptide-II-Induced Opening of Blood–Tumor Barrier. J Mol Neurosci 57, 11–20 (2015). https://doi.org/10.1007/s12031-015-0604-8

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