Metabolic responses to in vitro zinc supplementation

The present study was carried out to determine the effects and possible mechanism of action of zinc supplementation on the in vitro growth of malignant murine melanoma (B16) and non-malignant monkey kidney (LLCMK) cells. Cell culture studies showed that zinc supplementation significantly inhibited B16 growth at all the concentrations studied (1, 3, 5 and lOμg/ml). Zinc was also found to inhibit the growth of the LLCMK cells, although to a lesser extent than the B16 cells. Possible evidence of mobilisation of the essential fatty acids from the membrane phospholipid stores was noted in both cell types. This effect was, however, greater in the B16 cells. Δ⁶-desaturase activity was found to be significantly lower in the B16 cells than in the LLCMK cells (p ≥ 0.05). Zinc supplementation resulted in an increase in the enzymes activity in the LLCMK cells and, at high concentrations, in the B16 cells. An estimation of elongase and Δ⁶-desaturase activity with zinc supplementation indicated that zinc had little or no effect on the activity of these enzymes. B16 cells were found to have higher levels of free radicals than the LLCMK cells. Zinc supplementation resulted in increased free radical formation in the B16 cells, while no effect was observed in the LLCMK cells. Lipid peroxidation increased in both cell types with increased zinc concentrations. The observed effect of zinc supplementation on cell growth may involve these elevated levels of lipid peroxides. CycIo-oxygenase activity was found to be greater in the B16 cells than the LLCMK cells. The activity of the enzyme increased with higher concentrations of zinc (lOμg/ml) in both cell types. Prostaglandin E, levels were found to be lower in the B16 cells compared to the LLCMK cells. The levels of prostaglandin E, in both cell types appeared to be dependent on the levels of the polyunsaturated fatty acid precursors to the prostaglandins. Zinc was found to inhibit the activity of the enzyme adenylate cyclase in both cell types. The cAMP levels in the LLCMK cells were also found to decrease with zinc supplementation. In the case of the B16 cells, cAMP levels increased at low concentrations of zinc despite a decrease in adenyl ate cyclase activity, suggesting a possible inhibition of cAMP phosphodiesterase activity at these concentrations of zinc. It is concluded that although zinc supplementation does have an effect on cell growth, this effect is not mediated through the activation of adenylate cyclase by the prostaglandins resulting in elevated levels of cAMP. A possible mechanism involving lipid peroxidation is proposed