Abstract
Serum is a common supplement for cell culture due to it containing the essential active components for the growth and maintenance of cells. However, the knowledges of the active components in serum are incomplete. Apart from the direct influence of serum components on cultured cells, the reaction of serum components with tested drugs cannot be ignored, which usually results in the false conclusion on the activity of the tested drugs. Here we report the toxicity effect of polyamines (spermidine and spermine) on cultured cells, especially on drug-resistant cancer cell lines, which resulted from the oxidative degradation of polyamines by amine oxidases in serum supplement. Upon adding spermidine or spermine, high concentration of H2O2, an enzyme oxidation product of polyamines, was generated in culture media containing ruminant serum, such as fetal bovine serum (FBS), calf serum, bovine serum, goat serum or horse serum, but not in the media containing human serum. Drug-resistant cancer cell lines showed much higher sensitivity to the oxidation products of polyamines (H2O2 and acrolein) than their wild cell lines, which was due to their low antioxidative capacity.
Highlights
Cell culture is a widely used tool to study physiological, biological and pharmacological activities in vitro, as well as to produce biological components, such as proteins, hormones and vaccines
Since TETA, an unnatural compound with a structure similar to spermine, and putrescine are not the substrates of amine oxidases; above suggests that the cytotoxicity effects may not be caused by spermine and spermidine directly, but may be related to the metabolites of polyamines
We further measured the cytotoxicity effect of the enzymatic oxidation products of spermine, H2O2 and acrolein on the tested cell lines. Both of the two metabolites exhibited a dose-dependent toxicity effect on all cell lines, and the toxicity effect on drug-resistant cell lines was much stronger than that on wild cell lines (Fig. 1E,F). These results suggest that the cytotoxicity activity of spermine and spermidine may result from the enzymatic oxidation products of amine oxidases in fetal bovine serum (FBS)
Summary
The toxicity effect of polyamines on cultured cells. Polyamines (including spermine analogues) have been reported to have cytotoxicity effect on different cells[24,25,26,27]. The cytotoxicity assay of spermine and spermidine in the media containing 10% goat serum was performed Both polyamines exhibited higher toxicity effects to all the tested cell lines than those measured in the media containing 10% FBS, which was consistent with the higher concentrations of H2O2 generated in the media containing 10% goat serum (Fig. S5). The drug-resistant cell lines (A549T and MCF-7R) showed much higher sensitivity to spermine and spermidine than their wild cell lines (A549 and MCF-7) These results further confirm that it is the oxidation products of polyamines caused their cytotoxicity to cultured cells. The higher sensitivity of the drug-resistant cancer cells to the cytotoxicity of spermine and spermidine was demonstrated to be the lower antioxidative capacity of drug-resistant cells than wild cells, which caused higher oxidative stresses and resulted in the loss of ΔΨm and subsequent cell death. Before drawing a conclusion of the drug effect on cells, the side effect from the serum supplement should be excluded firstly
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