Arachidonate lipoxygenases (LOX) and their products play an important role in mediating growth factor-supported tumor cell proliferation and growth. The LOX pathway may also be critical in regulating tumor cell survival and apoptosis. Blocking the 12-LOX gene expression with sequence-specific antisense oligos or its activity with general or isoform-specific LOX inhibitors induces a strong apoptotic response in rat W256 carcinosarcoma cells of the monocytoid origin (Tang et al., 1996, Proc. Natl. Acad. Sci. U.S.A., 93:5241-5246). In the present study, several molecular approaches confirmed the predominant expression of platelet-type 12-LOX in W256 cells, with no or little expression of 5- and 15-LOX. NDGA, a general LOX inhibitor and BHPP, a 12-LOX-selective inhibitor, induced rapid and dose-dependent apoptosis of serum-cultured W256 cells as well as several other tumor (in particular leukemia) cell lines, thus suggesting a potential role for LOX in mediating serum-supported tumor cell survival. The molecular mechanism of NDGA-induced W256 cell death was subsequently investigated. NDGA-induced apoptosis could be significantly postponed by overexpression of 12-LOX, thus suggesting that the NDGA effect is, at least partly, dependent on its inhibition of LOX (i.e., 12-LOX). W256 cell apoptosis induced by NDGA could also be effectively inhibited by GSH-elevating or thiol agents as well as by lipid peroxidation inhibitors and an inhibitor of mitochondria respiratory chain rotenone. Further experiments demonstrated that NDGA treatment triggered rapid lipid peroxidation leading to the depletion of cytosolic and mitochondrial GSH pools. Interestingly, the lipid peroxidation induced by NDGA could not be inhibited by conventional free radical scavengers nor by cyclooxygenase or cytochrome P-450 monooxygenase inhibitors. In summary, the present work suggests a role of 12-LOX in regulating serum (growth factor)-supported survival of certain tumor cells.