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PMID: 11313437 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Blocking swelling-activated chloride current inhibits mouse liver cell proliferation.

The Journal of physiology ·Vol. 532 ·No. Pt 3 ·2001-05-01 ·Pages 661-72

Wondergem R, Gong W, Monen SH, Dooley SN, Gonce JL, Conner TD, Houser M, Ecay TW, Ferslew KE

Abstract

A non-transformed mouse liver cell line (AML12) was used to show that blocking swelling-activated membrane Cl- current inhibits hepatocyte proliferation. Two morphologically distinguishable cell populations exhibited distinctly different responses to hypotonic stress. Hypotonic stress (from 280 to 221 mosmol kg(-1)) to rounded, dividing cells activated an ATP-dependent, outwardly rectifying, whole-cell Cl- current, which took 10 min to reach maximum conductance. A similar anionic current was present spontaneously in 20 % of the dividing cells. Hypotonic stress to flattened, non-dividing cells activated no additional current. The Eisenman halide permeability sequence of swelling-activated anionic current in the dividing cells was SCN(-) > I(-) > Br(-) > Cl(-) > gluconate. Addition of either 4,4'-diisothiocyanatostilbene-2,2'-disulfonate (DIDS), 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB), tamoxifen or mibefradil inhibited swelling-activated anionic current. Hyperosmolarity by added sucrose inhibited the spontaneous anionic current in dividing cells. Added Cl- channel blockers NPPB (IC50 = 40 microM), DIDS (IC50 = 31 microM), tamoxifen (IC50 = 1.3 microM) and mibefradil (IC50 = 7 microM) inhibited proliferative growth of AML12 as determined by cell counts over 4 days or by protein accumulation over 2 days. Only the inhibitory effects of NPPB and mibefradil reversed with the drug washout. Hyperosmolarity by added sucrose (50 and 100 mM) also inhibited cell proliferation. Of the hydrophobic inhibitors neither NPPB at 40 microM nor tamoxifen at 1.3 microM, added for 48 h, reduced cellular ATP; however, DIDS at 31 microM significantly reduced cellular ATP with an equivalent increase in cellular ADP. We conclude that those membrane Cl- currents that can be activated by hypotonic stress are involved in mechanisms controlling liver cell growth, and that NPPB, tamoxifen and mibefradil at their IC50 for growth do not suppress the metabolism of mouse hepatocytes.

MeSH Terms
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid/pharmacology Adenosine Diphosphate/metabolism Adenosine Triphosphate/metabolism Animals Calcium Channel Blockers/pharmacology Carcinogens/pharmacology Cell Division/drug effects,physiology Cell Line Chloride Channels/antagonists & inhibitors,physiology Chlorides/metabolism Hepatocytes/cytology,physiology Hypertonic Solutions/pharmacology Liver/cytology Membrane Potentials/drug effects,physiology Mibefradil/pharmacology Mice Nitrobenzoates/pharmacology Patch-Clamp Techniques Sucrose/pharmacology Tamoxifen/pharmacology Water-Electrolyte Balance/drug effects,physiology
Chemicals
Calcium Channel Blockers Carcinogens Chloride Channels Chlorides Hypertonic Solutions Nitrobenzoates Tamoxifen Mibefradil 5-nitro-2-(3-phenylpropylamino)benzoic acid Sucrose Adenosine Diphosphate Adenosine Triphosphate 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Wondergem R
Department of Physiology, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN 37614-0576, USA. wonderge@etsu.edu
Gong W
Monen S H
Dooley S N
Gonce J L
Conner T D
Houser M
Ecay T W
Ferslew K E
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Article Info
Journal
The Journal of physiology
Abbr.
J Physiol
ISSN
0022-3751
DOI
10.1111/j.1469-7793.2001.0661e.x
Email
wonderge@etsu.edu
Published
2001-05-01
Pages
661-72
Language
English
Region
England
NLM ID
0266262
PMCID
PMC2278564
Subset
IM
Corrections
CommentIn
PMID: 11313429
External Links
PubMed source DOI: 10.1111/j.1469-7793.2001.0661e.x PMC Full Text View journal details
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