Ectophosphatase activity in the triple-negative breast cancer cell line MDA-MB-231.

Breast cancer is one of the most common cancers in the female population worldwide, and its development is thought to be associated with genetic mutations that lead to uncontrolled and accelerated growth of breast cells. This abnormal behavior requires extra energy, and indeed, tumor cells display a rewired energy metabolism compared to normal breast cells. Inorganic phosphate (Pi) is a glycolytic substrate of glyceraldehyde-3-phosphate dehydrogenase and has an important role in cancer cell proliferation. For cells to obtain Pi, ectoenzymes in the plasma membrane with their catalytic site facing the extracellular environment can hydrolyze phosphorylated molecules, and this is an initial and possibly limiting step for the uptake of Pi by carriers that behave as adjuvants in the process of energy harvesting and thus partially contributes to tumor energy requirements. In this study, the activity of an ectophosphatase in MDA-MB-231 cells was biochemically characterized, and the results showed that the activity of this enzyme was higher in the acidic pH range and that the enzyme had a Km = 4.5 ± 0.5 mM para-nitrophenylphosphate and a Vmax = 2280 ± 158 nM × h-1 × mg protein-1 . In addition, classical acid phosphatase inhibitors, including sodium orthovanadate, decreased enzymatic activity. Sodium orthovanadate was able to inhibit ectophosphatase activity while also inhibiting cell proliferation, adhesion, and migration, which are important processes in tumor progression, especially in metastatic breast cancer MDA-MB-231 cells that have higher ectophosphatase activity than MCF-7 and MCF-10 breast cells.

Inorganic phosphate transporters in cancer: Functions, molecular mechanisms and possible clinical applications.

Inorganic phosphate is one of the most essential nutrients for the maintenance of cell life. Because of its essential role in nutrient supplementation, the study of plasma membrane inorganic phosphate transporters in cancer biology has received much attention in recent years. Several studies suggest that these transporters are up-regulated in tumor cells and thus have been considered to be important promoters of tumor progression. Altered expression levels of inorganic phosphate transporters, such as NaPi-IIb (SLC34A2) and PiT-1 (SLC20A1), have been demonstrated. The purpose of this review article was to gather the relevant experimental records on inorganic phosphate transporters in tumors and to demonstrate the importance of these proteins in clinical applications. In this work, we demonstrate that for decades, the potential use of the inorganic phosphate transporter as an antigen for the diagnosis of tumor subtypes remained unknown. With the advancement in molecular biology techniques, phosphate transporters have been identified as being associated with cancer. In addition to their altered expression in cancer, several studies have demonstrated other functions of inorganic phosphate transporters, such as transceptors, rearrangements with oncogenes and modifications in the expression of ABC transporters, aiding in the process of proliferation and drug resistance.

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 NaPi-IIb carrier (SLC34A2), suggesting that this carrier may play a role in breast cancer progression. However, the biochemical behavior of inorganic phosphate (Pi) 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 Pi 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 Pi transport. We observed that the inorganic phosphate is dependent on sodium transport (K0,5 value = 21.98 mM for NaCl). Furthermore, the transport is modulated by different pH values and increasing concentrations of Pi, following the Michaelis-Menten kinetics (K0,5 = 0.08 mM Pi). PFA, monensin, furosemide and ouabain inhibited Pi transport, cell migration and adhesion. CONCLUSIONS: Taken together, these results showed that the uptake of Pi 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.

Venous thrombosis risk: effects of palm oil and hydrogenated fat diet in rats.

OBJECTIVE: We tested whether diets containing partially hydrogenated fat (PHVO, rich in trans fatty acids) or palm oil (PO, rich in saturated fat-C16 palmitic fatty acid) had different effects on the propensity for venous thrombosis, a marker of haemostatic cardiovascular risk. METHODS: Female Wistar rats were fed normolipidic diets containing PHVO or PO during lactation, and their young male pups were fed the same diets from weaning until the 180th day of life. We evaluated platelet fatty acid composition, serum lipids, platelet aggregation, clotting time, and venous thrombus formation. RESULTS: A significant and cumulative incorporation of trans fatty acid was observed only in the platelet lipids from the PHVO group, associated with an increased sensitivity to adenosine diphosphate (ADP) and venous thrombus formation in vivo. Platelets from rats raised on the PO diet also exhibited platelet aggregation induced by ADP and an increase in venous thrombus weight, with a concomitant increase in serum triglycerides. CONCLUSION: The prolonged replacement of dietary hydrogenated fat by PO impaired platelet aggregability and venous thrombosis, suggesting an increased risk of thromboembolic diseases.