Thus, the final net effects of 20-Rh2 and 20-Ppd did not exhibit remarkable P-gp regulation

containing glucose, none of the cells showed any decrease in cell 22564524 Catabolism of Exogenous Lactate in Breast Cancer viability or increase in cell death responses. Likewise, all cell lines treated with 0 20 mM in glucose-deprived conditions survived. Only at 40 mM lactate did MCF7 and MDA-MB-231 cells show significant cell death responses. Glucose deprivation usually elicits cell stress responses, such as activation of JNK1 and increased oxidative stress. This may suggest that very high lactate concentrations can augment the cellular stress elicited by glucose deprivation, but 40 mM lactate has been reported very infrequently in solid tumors and concentrations this high were not found in any of the breast tumor biopsies evaluated in this study. Thus, we conclude that cell viability is not appreciably affected over the range of physiologically relevant lactate concentrations observed in breast cancer with or without glucose. We AIC316 custom synthesis hypothesized that normal breast cells and breast cancer cells could utilize exogenous lactate for metabolic purposes. We used NMR with 13C-labeled lactate to track uptake; we first focused on the lactate metabolism in R3230Ac cells. In vitro, R3230Ac cells took up lactate in a concentration-dependent manner after 4 h of treatment. In glucosedeprived conditions, R3230Ac cells were treated with 5 mM 3-13C-lactate for 12 h. The 13C spectra of the cell lysate show peaks corresponding to 13C-lactate, 13C-alanine and 13C-glutamate. When these metabolites were normalized to protein levels, glutamate was the most abundant. To Catabolism of Exogenous Lactate in Breast Cancer and is regulated by the hypoxia inducible transcription factor, HIF-1. Previously, we have shown that cancer cell lines with high MCT1/low MCT4 expression consume more lactate than cancer cell lines with low MCT1/high MCT4 expression. Although it is well-documented that most normal cells and cancer cells typically express MCT1, MCT1 expression in R3230Ac cells has not been tested previously. R3230Ac tumor shows abundant membrane expression of MCT1, while MCT4 was undetectable, consistent with a lactateconsuming phenotype. It has previously been reported that R3230Ac cells utilize oxidative phosphorylation as well as glycolysis. Glutamate was found to be the predominant lactate-derived metabolite produced in R3230Ac cells in vitro. Glutamate is a TCA cycle by-product, formed from 1417961 a- ketoglutarate. This indicates that one pathway of lactate metabolism is respiration. In addition to glutamate formation indicating cellular respiration of lactate, R3230Ac cells were treated with increasing concentrations of lactate in vitro, and oxygen consumption rate was measured. All concentrations of lactate $ 10 mM tested showed a significant increase in oxygen consumption rate in the cells, providing further evidence that lactate is consumed via respiration. Kinetic uptake of glucose and lactate in vivo show lactate uptake in R3230Ac tumors occurs more rapidly than glucose uptake Kinetics of glucose and lactate uptake, retention, and clearance were measured using a novel scintillation probe following i.v. administration of either 14C-glucose or 14C-lactate. Catabolism of Exogenous Lactate in Breast Cancer A three-compartment pharmacokinetic model was formed from the data to determine rate constants for glucose and lactate uptake by the tumor and subcutaneous tissue. Rate constants for 14C-glucose and 14C-lactate are summarized in and pancreatic cancer, and that lactate containing glucose, none of the cells showed any decrease in cell Catabolism of Exogenous Lactate in Breast Cancer viability or increase in cell death responses. Likewise, all cell lines treated with 0 20 mM in glucose-deprived conditions survived. Only at 40 mM lactate did MCF7 and MDA-MB-231 cells show significant cell death responses. Glucose deprivation usually elicits cell stress responses, such as activation of JNK1 and increased oxidative stress. This may suggest that very high lactate concentrations can augment the cellular stress elicited by glucose deprivation, but 40 mM lactate has been reported 23570531 very infrequently in solid tumors and concentrations this high were not found in any of the breast tumor biopsies evaluated in this study. Thus, we conclude that cell viability is not appreciably affected over the range of physiologically relevant lactate concentrations observed in breast cancer with or without glucose. We hypothesized that normal breast cells and breast cancer cells could utilize exogenous lactate for metabolic purposes. We used NMR with 13C-labeled lactate to track uptake; we first focused on the lactate metabolism in R3230Ac cells. In vitro, R3230Ac cells took up lactate in a concentration-dependent manner after 4 h of treatment. In glucosedeprived conditions, R3230Ac cells were treated with 5 mM 3-13C-lactate for 12 h. The 13C spectra of the cell lysate show peaks corresponding to 13C-lactate, 13C-alanine and 13C-glutamate. When these metabolites were normalized to protein levels, glutamate was the most abundant. To Catabolism of Exogenous Lactate in Breast Cancer and is regulated by the hypoxia inducible transcription factor, HIF-1. Previously, we have shown that cancer cell lines with high MCT1/low MCT4 expression consume more lactate than cancer cell lines with low MCT1/high MCT4 expression. Although it is well-documented that most normal cells and cancer cells typically express MCT1, MCT1 expression in R3230Ac cells has not been tested previously. R3230Ac tumor shows abundant membrane expression of MCT1, while MCT4 was undetectable, consistent with a lactateconsuming phenotype. It has previously been reported that R3230Ac cells utilize oxidative phosphorylation as well as glycolysis. Glutamate was found to be the predominant lactate-derived metabolite produced in R3230Ac cells in vitro. Glutamate is a TCA cycle by-product, formed from a- ketoglutarate. This indicates that one pathway of lactate metabolism is respiration. In addition to glutamate formation indicating cellular respiration of lactate, R3230Ac cells were treated with increasing concentrations of lactate in vitro, and oxygen consumption rate was measured. All concentrations of lactate $ 10 mM tested showed a significant increase in oxygen consumption rate in the cells, providing further evidence that lactate is consumed via respiration. Kinetic uptake of glucose and lactate in vivo show lactate uptake in R3230Ac tumors occurs more rapidly than glucose uptake Kinetics of glucose and lactate uptake, retention, and clearance were measured using a novel scintillation probe following i.v. administration of either 14C-glucose or 14C-lactate. Catabolism of 18753409 Exogenous Lactate in Breast Cancer A three-compartment pharmacokinetic model was formed from the data to determine rate constants for glucose and lactate uptake by the tumor and subcutaneous tissue. Rate constants for 14C-glucose and 14C-lactate are summarized in and pancreatic cancer, and that lactate