Hydroquinone, a bioreactive metabolite of benzene, inhibits apoptosis in myeloblasts

Abstract

Hydroquinone (a major marrow metabolite of the leukemogen, benzene) induces incomplete granulocytic differentiation of mouse myeloblasts to the myelocyte stage, and also causes an increase in the number of myelocytes. This was confirmed using the normal interleukin 3 (IL-3)-dependent mouse myeloblastic 32D cell line. The hydroquinone-induced twofold increase in the number of IL-3-treated myelocytes does not result from stimulation of IL-3- induced proliferation. Hydroquinone's ability to effect this increase through an inhibition of apoptosis was investigated using mouse 32D and human HL-60 myeloblasts. Apoptosis induced by staurosporine treatment (0.5-1.0 μM) of HL-60 cells (50%) and 32D cells (15%) or by IL-3 withdrawal from 32D myeloblasts was determined by monitoring the development of characteristic morphological features and confirmed by the appearance of a typical nucleosomal DNA ladder upon agarose gel electrophoresis. Concentrations of hydroquinone (1-6 μM) that induce differentiation in 32D myeloblasts caused a concentration-dependent inhibition of staurosporine-induced apoptosis in both cell lines, with a 50% inhibitory concentration of 3 μM, and prevented apoptosis in IL-3-deprived 32D cells. Hydroquinone inhibition of apoptosis in myeloblasts, like hydroquinone-induced granulocytic differentiation, required myeloperoxidase-mediated oxidation of hydroquinone to its reactive species, p-benzoquinone, and was inhibited 50% by the peroxidase inhibitor, indomethacin (20 μM), p-benzoquinone (3 μM) was shown to cause a 50% inhibition of CPP32, an IL-1β-converting enzyme/Ced-3 cysteine protease involved in the implementation of apoptosis and present in myeloid cells. The ability of hydroquinone to induce a program of differentiation in the myeloblast that proceeds only to the myelocyte stage coupled with its ability to inhibit the CPP32 protease and, thereby, apoptosis of the proliferating myelocytes, may have important implications for benzene-induced acute myeloid leukemia.