Ca2+ retention in mitochondria, opening of the Cysclosporin A‐ sensitive permeability transition pore and cell death were studied in Ehrlich ascites tumour cells in the presence of different prooxidants. Low concentrations (1‐20 μM) of the prooxidants (menadione, cumenehydroperoxide, t‐butylhydroperoxide) induced pore‐opening in permeabilized cells at threshold Ca2+ load. Incubation of cells with low concentrations of prooxidants was able to induce cell cycle disturbance and cell death. Under the prooxidant effect, mitochondrial membrane potential drop and Ca2+ retention decrease in mitochondria were found to precede death of Ehrlich ascites tumour cells.