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pH regulation in HT29 colon carcinoma cells

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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 HCO −3 -containing and 7.21±0.09 (n=12) in HCO −3 -free solution. HOE-694 (10 μmol/l), a potent inhibitor of the Na+/H+ exchanger, did not affect control pHi. As a means to acidify cells we used the NH +4 /NH3 (20 mmol/l) prepulse technique. The mean peak acidification was 0.37±0.07 pH units (n=6). In HCC −3 -free solutions recovery from acid load was completely blocked by HOE-694 (1 μmol/l), whereas in HCO3 −3 -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 HCO −3 -containing and HCO −3 -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 HCO −3 -containing than in HCO −3 -free solutions. The present observations demonstrate three distinct mechanisms of pH regulation in HT29 cells: (a) a Na+/H+ exchanger, (b) a HCO −3 /Cl− exchanger and (c) a Na+-dependent HCC −3 transporter, probably the Na+-HCO −3 /Cl− antiporter. Under HCO −3 — free conditions the Na+/H+ exchanger fully accounts for recovery from acid load, whereas in HCO −3 -containing solutions this is accomplished by the Na+/H+ exchanger and a Na+-dependent mechanism, which imports HCO −3 . Recovery from alkaline load is caused by the HCO −3 /Cl− exchanger.

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Authors and Affiliations

  1. Physiologisches Institut, Albert-Ludwigs-Universität, Hermann-Herder-Strasse 7, D-79104, Freiburg, Germany

    M. Köttgen, J. Leipziger, K. -G. Fischer, R. Nitschke & R. Greger

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  1. M. Köttgen
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  2. J. Leipziger
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  3. K. -G. Fischer
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  4. R. Nitschke
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  5. R. Greger
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This study was supported by DFG Gr 480/10

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Köttgen, M., Leipziger, J., Fischer, K.G. et al. pH regulation in HT29 colon carcinoma cells. Pflügers Arch 428, 179–185 (1994). https://doi.org/10.1007/BF00374856

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  • DOI: https://doi.org/10.1007/BF00374856

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