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PMID: 7971175 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

pH regulation in HT29 colon carcinoma cells.

Pflugers Archiv : European journal of physiology ·Vol. 428 ·No. 2 ·1994-09-00 ·Pages 179-85

Köttgen M, Leipziger J, Fischer KG, Nitschke R, Greger R

Abstract

The pH regulation in HT29 colon carcinoma cells has been investigated using the pH-sensitive fluorescent indicator 2',7'-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF). Under control conditions, intracellular pH (pHi) was 7.21 +/- 0.07 (n = 22) in HCO3(-)-containing and 7.21 +/- 0.09 (n = 12) in HCO3(-)-free solution. HOE-694 (10 mumol/l), a potent inhibitor of the Na+/H+ exchanger, did not affect control pHi. As a means to acidify cells we used the NH4+/NH3 (20 mmol/l) prepulse technique. The mean peak acidification was 0.37 +/- 0.07 pH units (n = 6). In HCO3(-)-free solutions recovery from acid load was completely blocked by HOE-694 (1 mumol/l), whereas in HCO3(-)-containing solutions a combination of HOE-694 and 4,4'-diisothiocyanatostilbene-2,2'-disulphonate (DIDS, 0.5 mmol/l) was necessary to show the same effect. Recovery from acid load was Na(+)-dependent in HCO3(-)-containing and HCO3(-)-free solutions. Removal of external Cl- caused a rapid, DIDS-blockable alkalinization of 0.33 +/- 0.03 pH units (n = 15) and of 0.20 +/- 0.006 pH units (n = 5), when external Na+ was removed together with Cl-. This alkalinization was faster in HCO3(-)-containing than in HCO3(-)-free solutions. The present observations demonstrate three distinct mechanisms of pHi regulation in HT29 cells: (a) a Na+/H+ exchanger, (b) a HCO3-/Cl- exchanger and (c) a Na(+)-dependent HCO3- transporter, probably the Na(+)-HCO3-/Cl- antiporter.(ABSTRACT TRUNCATED AT 250 WORDS)

MeSH Terms
Acid-Base Equilibrium Antiporters/metabolism Bicarbonates/metabolism Chloride-Bicarbonate Antiporters Chlorides/metabolism Colonic Neoplasms/metabolism Fluorescence Guanidines/pharmacology Humans Hydrogen-Ion Concentration Intestinal Mucosa/metabolism Ion Transport Sodium/metabolism Sodium-Hydrogen Exchangers/antagonists & inhibitors,metabolism Sulfones/pharmacology Tumor Cells, Cultured
Chemicals
Antiporters Bicarbonates Chloride-Bicarbonate Antiporters Chlorides Guanidines Sodium-Hydrogen Exchangers Sulfones 3-methylsulfonyl-4-piperidinobenzoyl guanidine Sodium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Köttgen M
Physiologisches Institut, Albert-Ludwigs-Universität, Freiburg, Germany.
Leipziger J
Fischer K G
Nitschke R
Greger R
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Article Info
Journal
Pflugers Archiv : European journal of physiology
Abbr.
Pflugers Arch
ISSN
0031-6768
DOI
10.1007/BF00374856
Published
1994-09-00
Pages
179-85
Language
English
Region
Germany
NLM ID
0154720
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00374856 View journal details
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