Abstract
The feasibility of using a zinc-inducible gene expression system for the study of apoptosis-controlling genes in BAF-3 murine B cells and Jurkat human T cells was evaluated. Initially, cell sensitivity to a range of zinc concentrations was examined. It was found that zinc concentrations above 60 μM were toxic to BAF-3 cells and those above 50 μM were toxic to Jurkat cells. Secondly, the zinc concentration required to achieve maximal gene expression was examined. BAF-3 cells transiently transfected with the pMTCB6+/luciferase vector were exposed to zinc concentrations ranging from 0–120 μM, whilst stably transfected Jurkat cells were exposed to 0–70 μM zinc. At zinc concentrations nontoxic to each cell type, the maximum induction achieved was 20-fold (at 60 μM) in BAF-3 cells, and 7.5-fold (at 50 μM) in Jurkat cells. Thirdly, the effect of zinc on apoptosis was examined. It was shown that exposure to nontoxic zinc concentrations had no effect on IL-3 withdrawal-mediated apoptosis of BAF-3 cells. However, in the case of Jurkat cells, pre-exposure to zinc augmented CD95-mediated apoptosis. These results illustrate the importance of characterizing individual cell lines when using zinc-inducible gene expression systems.
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