Cellular Density and Cell Type are the Key Factors in Growth Inhibition Induced by 2,5Bis [1-aziridinyl]-1,4 benzoquinone (DZQ)

Abstract

Background: Cell density regulates the expression of various antioxidant enzymes in cell culture. The aim of this study was to study the effect of 2,5 bis-[1-aziridinyl]-1,4 benzoquinone (DZQ), an antitumor quinone bioactivated by NQO1, on HeLa and HepG2 cells cultured at various cell densities. Materials and Methods: Quinone toxicity was determined by a colorimetric growth inhibition assay. NQO1 and catalase activities were measured spectrophotometrically in soluble fractions, and NQO1 polypeptide was quantified by immunostaining with a commercial polyclonal antiserum. Results: As reported previously, NQO1 activity was much higher in confluent HeLa cells than in sparse cells. However, HepG2 cultures showed an opposite pattern in the regulation of this antioxidant enzyme, sparse cell cultures showing higher NQO1 activity similar to that found in confluent HeLa cells. The expression pattern of catalase activity was similar to that of NQO1 in HeLa cells, but this activity was constant and cell density-independent in HepG2. The growth inhibition effect of DZQ, correlated with NQO1 activity within a given cell type, but HepG2 was always much more sensitive to DZQ than HeLa cells, even under conditions where NQO1 activity was high in HeLa but low in HepG2. Conclusion: These results suggest that NQO1 activity is a major factor for DZQ bioactivation, but this enzyme is not likely the sole factor involved in the growth inhibition mediated by DZQ. Since part of the cytotoxic effect of DZQ is mediated by H₂O₂, other antioxidant enzymes, mainly catalase, could modulate the different growth inhibition found between HeLa and HepG2 cells. In confluent HeLa cells, the higher activity of NQO1 coincides with an increment of catalase activity, thus, reducing the oxidative stress produced by the H₂O₂ formed.