Antioxidants and doxorubicin supplementation to modulate CD14 expression and oxidative stress induced by vitamin D3 and seocalcitol in HL60 cells

Abstract

1α,25-dihydroxyvitamin D 3 (VD 3) and the EB1089 analog are well known for their roles in the modulation of proliferation and the differentiation of several malignant cells. In addition, VD 3 or EB1089 displayed a high disposal of oxidant features and the ability to cause release of reactive oxygen species (ROS). We attempted to enhance HL60 cell differentiation and to limit ROS generation, by the association of deltanoids with doxorubicin and the antioxidants catalase (CAT), superoxide dismutase (SOD) and N-acetyl cystein (NAC). Differentiation of HL60 cells into monocytic lineage was studied by expression of mRNA, protein CD14 and functional differentiation by the nitroblue tetrazolium assay. The 2',7'-dichlorodihydrofluorescein diacetate (H 2-DCFDA) dye allowed to evaluate in situ ROS generation. When associated with 0.1 nM EB1089, 15 nM doxorubicin induced an increase of differentiated cell percentage from 29% to 87% and did not affect VD 3-treated cells. The association with doxorubicin also induced a significant increase of ROS release (p<0.05) versus VD 3 and EB1089-treated cells. These results correspond to additivity of individual effects of doxorubicin and deltanoids. Antioxidant agents (10 nM NAC, 50 U/ml SOD or 2000 U/ml CAT) were associated with 10 nM VD 3 or 1 nM EB1089 for 72 h. Compared to VD 3 and EB1089 treatments, associations with antioxidants induced a slight increase of differentiated cells and a significant increase of CD14 mRNA. The highest differentiation effect occurred in the case of the EB1089-NAC association. Antioxidants induced a decrease (p<0.05) in ROS release generated by VD 3 or EB1089 near the level of untreated cells. Thus, antioxidant agents demonstrated a protective effect against VD 3 and EB1089 oxidative cytotoxicity and an enhancement of the monocyte differentiation. Combinations of antioxidants with deltanoids could dissociate the oxidative stress and differentiation.

The Critical Role of Redox Homeostasis in Shikonin-Induced HL-60 Cell Differentiation via Unique Modulation of the Nrf2/ARE Pathway

“…Pretreatments with the redox modulators separately prior to drug treatment were proved to be a reasonable approach according to literatures [30, 31]. In the study of Krance et al, VD 3 -treated cells incubated with diethyl maleate (a GSH depletor) followed by BSO had higher expression of differentiation marker CD11b than unexposed cells [8].…”

Radiosensitizing effect of oleanolic acid on tumor cells through the inhibition of GSH synthesis in vitro

“…Cellular antioxidants are important for the protection of cells against ROS and free radicals generated through endogenous or exogenous oxidative stress (6)(7)(8). Glutathione (GSH), the tripeptide thiol L-γ-glutamyl-L-cysteinyl-glycine, is one of the most abundant antioxidants in the cells with concentrations in the low millimolar range.…”

Ascorbic acid inhibits TPA-induced HL-60 cell differentiation by decreasing cellular H2O2 and ERK phosphorylation

“…However, oxidative stress can affect HL-60 cell differentiation. A previous study revealed that antioxidants, catalase, superoxide dismutase and N-acetyl cysteine increase the differentiation rate of vitamin D-treated HL-60 cells (24). However, compared with vitamin D-treated HL-60 cells, antioxidant inhibits cell differentiation in TPA-treated HL-60 cells (25).…”