The Journal of Membrane Biology

, Volume 245, Issue 2, pp 107–115 | Cite as

Volume-Activated Chloride Currents in Fetal Human Nasopharyngeal Epithelial Cells



Volume-activated chloride channels have been studied by us extensively in human nasopharyngeal carcinoma cells. However, the chloride channels in the counterpart of the carcinoma cells have not been investigated. In this study, volume-activated chloride currents (Icl,vol) were characterized in normal fetal human nasopharyngeal epithelial cells using the whole-cell patch-clamp technique. Under isotonic conditions, nasopharyngeal epithelial cells displayed only a weak background current. Exposure to 47% hypotonic solution activated a volume-sensitive current. The reversal potential of the current was close to the calculated equilibrium potential for Cl. The peak values of the hypotonicity-activated current at +80 mV ranged from 0.82 to 2.71 nA in 23 cells. Further analysis indicated that the density of the hypotonicity-activated current in most cells (18/23) was smaller than 60 pA/pF. Only five cells presented a current larger than 60 pA/pF. The hypotonicity-activated current was independent of the exogenous ATP. Chloride channel inhibitors ATP, tamoxifen and 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB), inhibited the current dramatically. The anion permeability of the hypotonicity-activated chloride channels was I > Br > Cl > gluconate. Unexpectedly, in isotonic conditions, ATP (10 mM) activated an inward-rectified current, which had not been observed in the nasopharyngeal carcinoma cells. These results suggest that, under hypotonic challenges, fetal human nasopharyngeal epithelial cells can produce Icl,vol, which might be involved in cell volume regulation.


Patch-clamp technique Regulation of ion transport by cell volume Ion channel/epithelial cell Epithelial chloride transport 



This work was supported by the National Natural Science Foundation of China (30771106, 30870567, 30871267, 90913020 and U0932004) and the Science & Technology Innovation Fund of Guangdong Medical College (STIF201102).


  1. Alison MR, Brittan M, Lovell MJ, Wright NA (2006) Markers of adult tissue-based stem cells. Handb Exp Pharmacol 174:185–227PubMedCrossRefGoogle Scholar
  2. Bond T, Basavappa S, Christensen M, Strange K (1999) ATP dependence of the ICl, swell channel varies with rate of cell swelling. Evidence for two modes of channel activation. J Gen Physiol 113:441–456PubMedCrossRefGoogle Scholar
  3. Browne LE, Jiang LH, North RA (2010) New structure enlivens interest in P2X receptors. Trends Pharmacol Sci 31:229–237PubMedCrossRefGoogle Scholar
  4. Chen L, Wang L, Zhu L, Nie S, Zhang J, Zhong P, Cai B, Luo H, Jacob TJ (2002) Cell cycle–dependent expression of volume-activated chloride currents in nasopharyngeal carcinoma cells. Am J Physiol Cell Physiol 283:C1313–C1323PubMedGoogle Scholar
  5. Chen LX, Zhu LY, Jacob TJ, Wang LW (2007) Roles of volume-activated Cl currents and regulatory volume decrease in the cell cycle and proliferation in nasopharyngeal carcinoma cells. Cell Prolif 40:253–267PubMedCrossRefGoogle Scholar
  6. Chen B, Jefferson DM, Cho WK (2010) Characterization of volume-activated chloride currents in regulatory volume decrease of human cholangiocyte. J Membr Biol 235:17–26PubMedCrossRefGoogle Scholar
  7. Chou CY, Shen MR, Wu SN (1995) Volume-sensitive chloride channels associated with human cervical carcinogenesis. Cancer Res 55:6077–6083PubMedGoogle Scholar
  8. De Paiva CS, Pflugfelder SC, Li DQ (2006) Cell size correlates with phenotype and proliferative capacity in human corneal epithelial cells. Stem Cells 24:368–375PubMedCrossRefGoogle Scholar
  9. Greene SB, Gunaratne PH, Hammond SM, Rosen JM (2010) A putative role for microRNA-205 in mammary epithelial cell progenitors. J Cell Sci 123:606–618PubMedCrossRefGoogle Scholar
  10. Harvey VL, Saul MW, Garner C, McDonald RL (2010) A role for the volume regulated anion channel in volume regulation in the murine CNS cell line, CAD. Acta Physiol (Oxf) 198:159–168CrossRefGoogle Scholar
  11. Inoue H, Mori S, Morishima S, Okada Y (2005) Volume-sensitive chloride channels in mouse cortical neurons: characterization and role in volume regulation. Eur J Neurosci 21:1648–1658PubMedCrossRefGoogle Scholar
  12. Inoue H, Takahashi N, Okada Y, Konishi M (2010) Volume-sensitive outwardly rectifying chloride channel in white adipocytes from normal and diabetic mice. Am J Physiol Cell Physiol 298:C900–C909PubMedCrossRefGoogle Scholar
  13. Izumi K, Inoki K, Fujimori Y, Marcelo CL, Feinberg SE (2009) Pharmacological retention of oral mucosa progenitor/stem cells. J Dent Res 88:1113–1118PubMedCrossRefGoogle Scholar
  14. Lazarenko RM, Kondrats’kyi AP, Pohoriela N, Shuba IM (2005) Alterations in ATP dependence of swelling-activated Cl current associated with neuroendocrine differentiation of LNCaP human prostate cancer epithelial cells [in Russian]. Fiziol Zh 51:57–66PubMedGoogle Scholar
  15. Mao J, Wang L, Fan A, Wang J, Xu B, Jacob TJ, Chen L (2007) Blockage of volume-activated chloride channels inhibits migration of nasopharyngeal carcinoma cells. Cell Physiol Biochem 19:249–258PubMedCrossRefGoogle Scholar
  16. Mao J, Chen L, Xu B, Wang L, Li H, Guo J, Li W, Nie S, Jacob TJ, Wang L (2008) Suppression of ClC-3 channel expression reduces migration of nasopharyngeal carcinoma cells. Biochem Pharmacol 75:1706–1716PubMedCrossRefGoogle Scholar
  17. Mao J, Xu B, Li H, Chen L, Jin X, Zhu J, Wang W, Zhu L, Zuo W, Chen W, Wang L (2010) Lack of association between stretch-activated and volume-activated Cl currents in hepatocellular carcinoma cells. J Cell Physiol 226:1176–1185CrossRefGoogle Scholar
  18. Morris AP (1999) The regulation of epithelial cell cAMP- and calcium-dependent chloride channels. Adv Pharmacol 46:209–251PubMedCrossRefGoogle Scholar
  19. Okada Y (2006) Cell volume–sensitive chloride channels: phenotypic properties and molecular identity. Contrib Nephrol 152:9–24PubMedCrossRefGoogle Scholar
  20. Okada Y, Sato K, Numata T (2009) Pathophysiology and puzzles of the volume-sensitive outwardly rectifying anion channel. J Physiol 587:2141–2149PubMedGoogle Scholar
  21. Poletto Chaves LA, Varanda WA (2008) Volume-activated chloride channels in mice Leydig cells. Pflugers Arch 457:493–504PubMedCrossRefGoogle Scholar
  22. Qian JS, Pang RP, Zhu KS, Liu DY, Li ZR, Deng CY, Wang SM (2009) Static pressure promotes rat aortic smooth muscle cell proliferation via upregulation of volume-regulated chloride channel. Cell Physiol Biochem 24:461–470PubMedCrossRefGoogle Scholar
  23. Stutzin A, Hoffmann EK (2006) Swelling-activated ion channels: functional regulation in cell-swelling, proliferation and apoptosis. Acta Physiol (Oxf) 187:27–42CrossRefGoogle Scholar
  24. Sun XR, Chen LX, Mao JW, Zhu LY, Nie SH, Zhong P, Li P, Wang LW (2005a) Regulatory volume decrease and its mechanism in nasopharyngeal epithelial cells [in Chinese]. Shi Yan Sheng Wu Xue Bao 38:353–358PubMedGoogle Scholar
  25. Sun XR, Wang LW, Mao JW, Zhu LY, Nie SH, Zhong P, Chen LX (2005b) Background chloride currents in fetal human nasopharyngeal epithelial cells [in Chinese]. Sheng Li Xue Bao 57:349–354PubMedGoogle Scholar
  26. Verkman AS, Galietta LJ (2009) Chloride channels as drug targets. Nat Rev Drug Discov 8:153–171PubMedCrossRefGoogle Scholar
  27. Voets T, Wei L, De Smet P, Van Driessche W, Eggermont J, Droogmans G, Nilius B (1997) Downregulation of volume-activated Cl currents during muscle differentiation. Am J Physiol Cell Physiol 272:C667–C674Google Scholar
  28. Volk KA, Zhang C, Husted RF, Stokes JB (1996) Cl current in IMCD cells activated by hypotonicity: time course, ATP dependence, and inhibitors. Am J Physiol Renal Physiol 271:F552–F559Google Scholar
  29. Wang LW, Chen LX, Jacob T (2004) ClC-3 expression in the cell cycle of nasopharyngeal carcinoma cells. Sheng Li Xue Bao 56:230–236PubMedGoogle Scholar
  30. Yang L, Ye D, Ye W, Jiao C, Zhu L, Mao J, Jacob TJ, Wang L, Chen L (2011) ClC-3 is a main component of background chloride channels activated under isotonic conditions by autocrine ATP in nasopharyngeal carcinoma cells. J Cell Physiol 226:2516–2526PubMedCrossRefGoogle Scholar
  31. Zuo W, Zhu L, Bai Z, Zhang H, Mao J, Chen L, Wang L (2009) Chloride channels involve in hydrogen peroxide-induced apoptosis of PC12 cells. Biochem Biophys Res Commun 387:666–670PubMedCrossRefGoogle Scholar

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© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Institute of Aging Research, Key Laboratory for Medical Molecular Diagnostics of Guangdong ProvinceGuangdong Medical CollegeDongguanChina
  2. 2.Department of Pharmacology, Medical CollegeJinan UniversityGuangzhouChina
  3. 3.Department of PhysiologyGuangdong Medical CollegeZhanjiangChina
  4. 4.Department of Biology, Guangdong Key Laboratory for Bioactive Drugs ResearchGuangdong Pharmaceutical UniversityGuangzhouChina
  5. 5.Department of Physiology, Medical CollegeJinan UniversityGuangzhouChina

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