Characterization of inorganic phosphate transport in the triple-negative breast cancer cell line, MDA-MB-231

BACKGROUND: Recent studies demonstrate that interstitial inorganic phosphate is significantly elevated in the breast cancer microenvironment as compared to normal tissue. In addition it has been shown that breast cancer cells express high levels of the NaP(i)-IIb carrier (SLC34A2), suggesting that this carrier may play a role in breast cancer progression. However, the biochemical behavior of inorganic phosphate (P(i)) transporter in this cancer type remains elusive. METHODS: In this work, we characterize the kinetic parameters of Pi transport in the aggressive human breast cancer cell line, MDA-MB-231, and correlated P(i) transport with cell migration and adhesion. RESULTS: We determined the influence of sodium concentration, pH, metabolic inhibitors, as well as the affinity for inorganic phosphate in P(i) transport. We observed that the inorganic phosphate is dependent on sodium transport (K(0,5) value = 21.98 mM for NaCl). Furthermore, the transport is modulated by different pH values and increasing concentrations of P(i), following the Michaelis-Menten kinetics (K(0,5) = 0.08 mM P(i)). PFA, monensin, furosemide and ouabain inhibited P(i) transport, cell migration and adhesion. CONCLUSIONS: Taken together, these results showed that the uptake of P(i) in MDA-MB-231 cells is modulated by sodium and by regulatory mechanisms of intracellular sodium gradient. General Significance: Pi transport might be regarded as a potential target for therapy against tumor progression.