Signalling steps in apoptosis by ether lipids

Abstract

We have investigated the mechanisms of induction of apoptosis by the antineoplastic ether lipid ET-18-OCH₃ (ALP) in sensitive S49^wt mouse lymphoma cells and ALP-resistant S49^ar variants, both with wild-type p53, and in related L1210 cells with mutated p53. Ether lipid-resistant S49^ar cells were cross-resistant to extracellular stress factors (cold shock, heat shock, H₂O₂, dimethylsulfoxide) and to radiation-induced apoptosis but not to physiological apoptotic signals (dexamethasone, growth factor deprivation, thapsigargin, C₂-ceramide) and expressed similar levels of the apoptosis-regulating proteins Bcl-2, Bcl-X, Bax, Bad and Bak as did the parent S49^wt cells. The uptake of [³H]-ALP was strongly reduced in the stress-resistant cells but this was not associated with significant differences in membrane cholesterol:phospholipid content nor in membrane microviscosity. In S49^ar cells the activity of the antioxidant enzyme glutathione peroxidase (GSH-Px) was increased 4-fold and depletion of glutathione with the drug L-buthionine-S-R-sulfoximine (L-BSO) lowered the resistance of S49^ar cells to ALP, stress factors and ionising radiation. The results indicate that ether lipids induce apoptosis by imposing a special form of physico-chemical stress, mediated by reactive oxygen species but independent of p53 status. The capacity of glutathione-dependent anti-oxidant defence appeared an important and shared determinant of the sensitivity to ether lipids, several types of extracellular stress and ionising radiation.