Antitumor activity of nigericin and 5-(N-ethyl-N-isopropyl)amiloride: an approach to therapy based on cellular acidification and the inhibition of regulation of intracellular pH.
The extracellular pH (pHe) in solid tumors is frequently lower than the pHe in normal tissues, but the intracellular pH (pHi) is regulated to physiological levels. Cell killing can be achieved in an acidic environment in tissue culture by nigericin, which acidifies cells by transporting H+ from the extracellular space into the cytoplasm; this cell killing can be enhanced when used with 5-(N-ethyl-N-isopropyl)amiloride (EIPA), a potent inhibitor of membrane-based Na+/H+ exchange, which plays a major role in the regulation of pHi (R. P. Maidorn; E. J. Cragoe; I. F. Tannock, Br. J. Cancer 67:297-303; 1993). We have therefore assessed the ability of nigericin and EIPA to kill cells in two murine solid tumors (the KHT fibrosarcoma and the EMT-6 sarcoma). Hydralazine, which reduces tumor blood flow, or glucose, which stimulates glycolysis leading to accumulation of lactate, were also administered to mice to lower pHe in the tumors. We observed only a small decrease in the surviving fractions of cells in the tumors when tolerated doses of nigericin and EIPA were given IP to tumor-bearing mice. When nigericin and EIPA were combined with administration of hydralazine, the surviving fraction of cells in both tumors was reduced by a factor of 0.01, but there were minimal effects on growth delay. Administration of glucose with nigericin and EIPA led to a smaller reduction in surviving fraction of the KHT tumor (by approximately 0.1), although glucose was more effective than hydralazine in lowering the mean tumor pHe. When KHT tumors were treated with 15 Gy X-rays followed immediately by nigericin, EIPA, and hydralazine, a reduced surviving fraction as well as an increase in tumor growth delay was observed compared to radiation alone; however, there was little evidence to suggest that these agents were selectively toxic to the cells that survived radiation. Nigericin and EIPA, with or without hydralazine, had minimal effects on normal tissues, as assessed by changes in body weight, number of leukocytes, and serum creatinine levels. We conclude that pharmacological effects to acidify cells and to prevent regulation of pHi under the acidic conditions that exist in solid tumors can lead to moderate levels of cell killing, if additional strategies are used to lower tumor pHe.
Hasuda
K
K
Department of Medicine, Ontario Cancer Institute, University of Toronto, Canada.
Lee
C
C
Tannock
I F
IF
D000584:Amiloride / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D000818:Animals; D015687:Cell Hypoxia; D002470:Cell Survival / Q000187:drug effects; D003131:Combined Modality Therapy; D004354:Drug Screening Assays, Antitumor; D004357:Drug Synergism; D005260:Female; D005354:Fibrosarcoma / Q000737:chemistry / Q000188:drug therapy* / Q000473:pathology / Q000532:radiotherapy; D005947:Glucose / Q000627:therapeutic use; D006830:Hydralazine / Q000627:therapeutic use; D006863:Hydrogen-Ion Concentration / Q000187:drug effects*; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D009368:Neoplasm Transplantation; D009550:Nigericin / Q000494:pharmacology / Q000627:therapeutic use*; D009940:Organoids / Q000187:drug effects / Q000473:pathology; D011522:Protons; D012513:Sarcoma, Experimental / Q000737:chemistry / Q000188:drug therapy* / Q000473:pathology / Q000532:radiotherapy
D011522:Protons; D006830:Hydralazine; D000584:Amiloride; D005947:Glucose; D009550:Nigericin; C039614:ethylisopropylamiloride