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1 Department of Physiology and Biophysics, Seoul National University College of Medicine, Seoul, Korea
2 Department of Aesthesiology, Sungkyunkwan University School of Medicine, Suwon, Korea
3 Department of Microbiology and Institute of Biomedical Science, Hanyang University College of Medicine, Seoul, Korea
4 Department of Internal Medicine, Seoul National University College of Medicine, Liver Research Institute, Seoul, Korea
* To whom correspondence should be addressed. E-mail: Insuk{at}plaza.snu.ac.kr.
Volume regulation is essential for cell function, but it is unknown which channels are involved in a regulatory volume decrease (RVD) in human gastric epithelial cells. Exposure to a hypotonic solution caused the increase in AGS cell volume followed by the activation of a current. The reversal potential of the swelling-induced current suggested that Cl- was the primary charge carrier. The selectivity sequence for different anions was I- > Br- > Cl- > F- > gluconate. This current was inhibited by flufenanmate, 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS), tamoxifen and 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB). Intracellular dialysis of three different anti-ClC-3 antibodies abolished or attenuated the Cl- current and disrupted RVD, whereas the current and RVD was unaltered by anti-ClC-2 antibody. Immunoblot studies demonstrated the presence of ClC-3 protein in Hela and AGS cells. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis detected expression of ClC-3, MDR-1 and pICln mRNA in AGS cells. These results suggest a fundamental role of endogenous ClC-3 in the swelling-activated Cl- channels function and cell volume regulation in human gastric epithelial cells.
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