Thymoquinone effect on the Dictyostelium discoideum model correlates with functional roles for glutathione S-transferases in eukaryotic proliferation, chemotaxis, and development

An increasing body of literature demonstrates the therapeutic relevance of polyphenols in eukaryotic cell and animal model studies. The phase II glutathione S-transferases (GST) show differential responses to thymoquinone, a major bioactive polyphenol constituent of the black seed, Nigella sativa. Beyond antioxidant defense, GSTs may act in non-enzymatic capacities to effect cell cycle, motility, and differentiation. Here, we report the impact of thymoquinone on the life cycle of the eukaryotic model Dictyostelium discoideum and accompanying profiles of its GST-alpha (DdGSTA) enzyme activity and isozyme expression. In silico molecular modeling revealed strong interaction(s) between thymoquinone and DdGSTA2 and DdGSTA3 isozymes that correlated with in vivo, dose-dependent inhibition of cell proliferation of amoebae at 24, 48, and 72hr. Similarly, cytosolic DdGST enzyme activity (CDNB activity) was also responsive to different thymoquinone concentrations. Thymoquinone generally reduced expression of DdGSTA2 and DdGSTA3 isozymes in proliferating cells, however differential expression of the isozymes occurred during starvation. Thymoquinone effectively reduced early-stage aggregation of starved amoeba, accompanied by increased reactive oxygen species and altered expression of tubulin and contact site A (gp80), which resulted in reduced morphogenesis and fruiting body formation. These observations reveal that thymoquinone can impact signaling mechanisms that regulate proliferation and development in D. discoideum.