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Altered cytotoxicity of ROS-inducing compounds by sodium pyruvate in cell culture medium depends on the location of ROS generation.

Kelts JL, Cali JJ, Duellman SJ, Shultz J - Springerplus (2015)

Bottom Line: Three compounds, eseroline, benserazide, and pyrogallol induced H2O2 in cell culture media independent of cells.However, another compound, menadione, induced H2O2 in a manner largely dependent on the MDA-MB-231 breast cancer cells used in this study, which is consistent with its known mechanism of inducing ROS through intracellular redox cycling. 1 mM pyruvate, as well as catalase, reduced the H2O2 in culture wells with each ROS inducer tested but it only reduced the cytotoxicity of cell-independent inducers.It reduced the cytotoxicity of benserazide and pyrogallol >10-fold and of eseroline about 2.5-fold, but had no effect on menadione cytotoxicity.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry and Biochemistry, University of Michigan-Flint, 556 Murchie Science Building 303 E. Kearsley St., Flint, MI 48502 USA.

ABSTRACT
Induction of oxidative stress by drugs and other xenobiotics is an important mechanism of cytotoxicity. However, in vitro studies on the relationship between oxidative stress and cytotoxicity in cultured cells is frequently complicated by the fact that cell culture medium components affect reactive oxygen species (ROS) exposures in ways that vary with the mode of ROS production. The objectives of this study were to first determine the mode of ROS induction by certain model compounds when they are applied to cultured cells, and then to determine how ROS induction and cytotoxicity were affected by the ROS-quenching medium component pyruvate. Three compounds, eseroline, benserazide, and pyrogallol induced H2O2 in cell culture media independent of cells. However, another compound, menadione, induced H2O2 in a manner largely dependent on the MDA-MB-231 breast cancer cells used in this study, which is consistent with its known mechanism of inducing ROS through intracellular redox cycling. 1 mM pyruvate, as well as catalase, reduced the H2O2 in culture wells with each ROS inducer tested but it only reduced the cytotoxicity of cell-independent inducers. It reduced the cytotoxicity of benserazide and pyrogallol >10-fold and of eseroline about 2.5-fold, but had no effect on menadione cytotoxicity. From this data, it was concluded that depending on the mechanism of ROS induction, whether intra- or extracellular, a ROS-quenching medium component such as pyruvate will differentially affect the net ROS-induction and cytotoxicity of a test compound.

No MeSH data available.


Related in: MedlinePlus

Effect of sodium pyruvate on cytotoxicty. MDA-MB-231 cells were incubated with increasing amounts of test compounds a alamethicin, b menadione c eseroline, d benserazide, or e pyrogallol for 68 h. Cell viability was determined by luminescent ATP detection. Results are shown as percent viable cells, mean ± SEM. Data is the average of three separate experiments with quadruplicate determinations of each point. Filled square MEM, filled circle MEM plus 1 mM pyruvate.
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Fig4: Effect of sodium pyruvate on cytotoxicty. MDA-MB-231 cells were incubated with increasing amounts of test compounds a alamethicin, b menadione c eseroline, d benserazide, or e pyrogallol for 68 h. Cell viability was determined by luminescent ATP detection. Results are shown as percent viable cells, mean ± SEM. Data is the average of three separate experiments with quadruplicate determinations of each point. Filled square MEM, filled circle MEM plus 1 mM pyruvate.

Mentions: To examine the cytotoxicity of our test compounds and the impact of extracellular H2O2 quenching we measured cell viability after exposing MDA-MB-231 cells in MEM to a range of compound concentrations in the presence or absence of 1 mM sodium pyruvate. Pyruvate had minimal or no impact on the toxicity of alamethacin and menadione, which do not produce substantial ROS in MEM without cells (Figure 4). EC50 values express the toxic potency of each compound (Table 1).Figure 4


Altered cytotoxicity of ROS-inducing compounds by sodium pyruvate in cell culture medium depends on the location of ROS generation.

Kelts JL, Cali JJ, Duellman SJ, Shultz J - Springerplus (2015)

Effect of sodium pyruvate on cytotoxicty. MDA-MB-231 cells were incubated with increasing amounts of test compounds a alamethicin, b menadione c eseroline, d benserazide, or e pyrogallol for 68 h. Cell viability was determined by luminescent ATP detection. Results are shown as percent viable cells, mean ± SEM. Data is the average of three separate experiments with quadruplicate determinations of each point. Filled square MEM, filled circle MEM plus 1 mM pyruvate.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4469600&req=5

Fig4: Effect of sodium pyruvate on cytotoxicty. MDA-MB-231 cells were incubated with increasing amounts of test compounds a alamethicin, b menadione c eseroline, d benserazide, or e pyrogallol for 68 h. Cell viability was determined by luminescent ATP detection. Results are shown as percent viable cells, mean ± SEM. Data is the average of three separate experiments with quadruplicate determinations of each point. Filled square MEM, filled circle MEM plus 1 mM pyruvate.
Mentions: To examine the cytotoxicity of our test compounds and the impact of extracellular H2O2 quenching we measured cell viability after exposing MDA-MB-231 cells in MEM to a range of compound concentrations in the presence or absence of 1 mM sodium pyruvate. Pyruvate had minimal or no impact on the toxicity of alamethacin and menadione, which do not produce substantial ROS in MEM without cells (Figure 4). EC50 values express the toxic potency of each compound (Table 1).Figure 4

Bottom Line: Three compounds, eseroline, benserazide, and pyrogallol induced H2O2 in cell culture media independent of cells.However, another compound, menadione, induced H2O2 in a manner largely dependent on the MDA-MB-231 breast cancer cells used in this study, which is consistent with its known mechanism of inducing ROS through intracellular redox cycling. 1 mM pyruvate, as well as catalase, reduced the H2O2 in culture wells with each ROS inducer tested but it only reduced the cytotoxicity of cell-independent inducers.It reduced the cytotoxicity of benserazide and pyrogallol >10-fold and of eseroline about 2.5-fold, but had no effect on menadione cytotoxicity.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry and Biochemistry, University of Michigan-Flint, 556 Murchie Science Building 303 E. Kearsley St., Flint, MI 48502 USA.

ABSTRACT
Induction of oxidative stress by drugs and other xenobiotics is an important mechanism of cytotoxicity. However, in vitro studies on the relationship between oxidative stress and cytotoxicity in cultured cells is frequently complicated by the fact that cell culture medium components affect reactive oxygen species (ROS) exposures in ways that vary with the mode of ROS production. The objectives of this study were to first determine the mode of ROS induction by certain model compounds when they are applied to cultured cells, and then to determine how ROS induction and cytotoxicity were affected by the ROS-quenching medium component pyruvate. Three compounds, eseroline, benserazide, and pyrogallol induced H2O2 in cell culture media independent of cells. However, another compound, menadione, induced H2O2 in a manner largely dependent on the MDA-MB-231 breast cancer cells used in this study, which is consistent with its known mechanism of inducing ROS through intracellular redox cycling. 1 mM pyruvate, as well as catalase, reduced the H2O2 in culture wells with each ROS inducer tested but it only reduced the cytotoxicity of cell-independent inducers. It reduced the cytotoxicity of benserazide and pyrogallol >10-fold and of eseroline about 2.5-fold, but had no effect on menadione cytotoxicity. From this data, it was concluded that depending on the mechanism of ROS induction, whether intra- or extracellular, a ROS-quenching medium component such as pyruvate will differentially affect the net ROS-induction and cytotoxicity of a test compound.

No MeSH data available.


Related in: MedlinePlus