When Saccharomyces cerevisiae aerobic growth index cells are treated with acute dithione, they lose cell wall and membrane integrity and die from necrosis, and this behavior is related to the reduction of protein and non-protein sulfhydryl groups. A large increase in cellular reactive oxygen species (ROS) associated with mitochondrial membrane potential modification in cells treated with Dithianon was also detected. In filamentous fungi, Dithianon inhibits mycelial growth and conidial germination. Studies on Ehrlich ascites carcinoma and yeast cells have shown that Dithianon inhibits respiration and fermentation and affects several thiol enzymes in the glycolysis pathway, such as hexokinase, phosphofructokinase, and glycoeraldeyde-3-phosphate dehydrogenase. Dithianon has cytotoxicity in vitro and affects the cell transformation activity of BLAB/c 3 T3 cells.

The activity of testosterone hydroxylase in liver microsomes of male mice increases after acute doses of Dithianon treatment, while female mice show inactivation.