Effect of NYD-SP27 down-regulation on ATP-induced Ca2+-dependent pancreatic duct anion secretion in cystic fibrosis cells

Authors

  • Jin Xia Zhu,

    1. Epithelial Cell Biology Research Center, Department of Physiology, Faculty of Medicine, Rm 410, Basic Medical Science Building, The Chinese University of Hong Kong, Shatin, Hong Kong, China
    2. Department of Physiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China
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  • Ning Yang,

    1. Department of Physiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China
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  • Hu Zhu,

    1. Epithelial Cell Biology Research Center, Department of Physiology, Faculty of Medicine, Rm 410, Basic Medical Science Building, The Chinese University of Hong Kong, Shatin, Hong Kong, China
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  • Yiu Wa Chung,

    1. Epithelial Cell Biology Research Center, Department of Physiology, Faculty of Medicine, Rm 410, Basic Medical Science Building, The Chinese University of Hong Kong, Shatin, Hong Kong, China
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  • Hsiao Chang Chan

    Corresponding author
    1. Epithelial Cell Biology Research Center, Department of Physiology, Faculty of Medicine, Rm 410, Basic Medical Science Building, The Chinese University of Hong Kong, Shatin, Hong Kong, China
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Corresponding author. Tel.: +852 2609 6839; fax: +852 2603 5022. hsiaocchan@cuhk.edu.hk

Abstract

Our previous study demonstrated that NYD-SP27 is a novel inhibitory PLC isoform expressed endogenously in human pancreas and upregulated in CFPAC-1 cells. The present study investigated the effect of NYD-SP27 down-regulation on the ATP-stimulated and Ca2+-dpendent pancreatic anion secretion by CFPAC-1 cell line using short-circuit current (I SC) recording. NYD-SP27 antisense-transfected CFPAC-1 (AT-CF) cells exhibited a significantly higher basal transmembrane potential difference and current than those of empty vector-transfected CFPAC-1 (VT-CF) cells. Cl channel blocker, DPC or Glibenclamide (1 mM), and inhibitor of Na+-K+-Cl cotransporter, bumetanide (100 μM) significantly inhibited the basal current in AT-CF cells. The inhibitor of adenylate cyclase, MDL12330A (20 μM), and Ca2+-dependent Cl channel (CaCC) blocker, DIDS (100 μM) also significantly reduced the basal current in AT-CF. Apical application of ATP (10 μM) stimulated a fast transient I SC increase in VT-CF cells, but a more sustained rise with slower decline in AT-CF cells. Pretreatment with BAPTA-AM (50 μM) reduced the ATP-induced I SC response in AT-CF cells by 77.9%. PMA (1 μM), a PKC activator, inhibited the ATP-stimulated current increase (the transient peak) in VT-CF cells, but had no effect on the AT-CF cells. However, PKC inhibitor, staurosporine (40 μM) could inhibit the ATP-induced I SC response in AT-CF cells. The present results confirm the previously proposed inhibitory role of NYD-SP27 in the PLC pathway and demonstrate that the suppression of its expression could result in an enhancement of ATP-stimulated Ca2+ dependent pancreatic anion secretion.

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