Inhibition of intracellular pH control and relationship to cytotoxicity of chlorambucil and vinblastine.
The uptake and cytotoxicity of weakly acidic or basic chemotherapeutic agents is determined in part by passive diffusion along the pH gradient between the intracellular and extracellular compartments. In vivo it is known that tumour extracellular pH is more acidic than intracellular pH. Using CaNT murine tumour cells in vitro, we found the cytotoxicity of chlorambucil (a weak acid) increased as the extracellular pH of the culture medium (pHmed) was acidified. The cytotoxicity of vinblastine shows a reverse pH relationship with reduced cytotoxicity as pHmed was acidified. Chlorambucil cytotoxicity increased at acidic pHmed because the weak acidic function is ionised to a lesser extent at acidic pH and, therefore, favours drug uptake into the relatively neutral intracellular compartment. Vinblastine cytotoxicity decreased at acidic pHmed because the weak basic function is ionised to a greater extent at acidic pH and therefore does not favour drug uptake into the relatively neutral intracellular compartment. Using a combination of an inhibitor of the cell membrane proton pump, amiloride, and the ionophore, nigericin, the intracellular compartment can be acidified. This results in a time-dependent increase in sensitivity of the cells to low pHmed with significant cytotoxicity after 6 h exposure to pHmed = 6.2 and suggests that there is potential for direct tumour cytotoxicity in vivo if the tumour extracellular pH were equally acidic. An indirect effect of intracellular acidification is to alter the distribution of drugs between the extra- and intracellular compartment by reducing the pH gradient across the cell membrane. In response to intracellular acidification, the cytotoxicity of chlorambucil was reduced and that for vinblastine was increased. Inhibition of cellular pH control may result in direct cytotoxicity by acidification due to inhibition of proton efflux or indirectly by resulting in differential uptake of chemotherapeutic agents with weak acidic or basic functions.
Parkins
C S
CS
CRC Tumour Microcirculation Group, Gray Laboratory Cancer Research Trust, Mount Vernon Hospital, Northwood, Middlesex, UK.
Chadwick
J A
JA
Chaplin
D J
DJ
D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D002699:Chlorambucil / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D051379:Mice; D009550:Nigericin / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014747:Vinblastine / Q000494:pharmacology*
D018906:Antineoplastic Agents, Alkylating; D000972:Antineoplastic Agents, Phytogenic; D002699:Chlorambucil; D014747:Vinblastine; D000584:Amiloride; D009550:Nigericin