Effect of Acidosis-Induced Signalling Pathways on Mitochondrial O2 Consumption of Tumour Cells.

Abstract

Signalling pathways such as ERK1/2, p38 or PI3K are activated in tumour cells by extracellular acidosis, which is a common phenomenon in human tumours. These signalling pathways can modulate the mitochondrial function and activity. The aim of the study was to evaluate the impact of extracellular acidosis on the mitochondrial O2 consumption and, in consequence, the potential role of ERK1/2, p38 and PI3K cascades on modulating the respiratory function. The O2 consumption rate (OCR) was measured at pH7.4 and extracellular acidosis (pH6.6) in combination with inhibition of the respective signalling pathway. The activity of the pathways was determined by phosphorylation-specific western blot using the cytosolic and mitochondrial fraction of cell lysates. The experiments were performed on a rat tumour cell line (subline AT1 of the rat R-3327 prostate carcinoma) and normal cells (NRK-49F fibroblasts). Acidosis increased the OCR of AT1 cells, especially the basal OCR and the O2 consumption, which is related to ATP production. In normal NRKF cells OCR was unaffected by low pH. Inhibition of ERK1/2 and PI3K, but not p38, reduced the acidosis-induced increase of the OCR significantly in AT1 tumour cells. In this cell line acidosis also led to an ERK1/2 and PI3K activation, predominantly in the mitochondrial fraction. These results indicate that extracellular acidosis activates cellular respiration in tumour cells, presumably by activating the ERK1/2 and/or the PI3K signalling cascade. This activation of ERK1/2 and PI3K is located primarily in the mitochondrial compartment of the cells.