4-Hydroxynonenal (4HNE) is the most prevalent toxic lipid peroxidation product formed during oxidative stress. It exerts its cytotoxicity mainly by the modification of intracellular
proteins. The detection of 4HNE-modified
proteins in several degenerative disorders suggests a role for 4HNE in the onset of these diseases. Efficient protection mechanisms are required to prevent the intracellular accumulation of 4HNE. The toxicity of 4HNE was tested with the
small cell lung cancer cell lines
GLC(4) and the multidrug-resistance-
protein (MRP1)-overexpressing counterpart
GLC(4)/Adr. In the presence of the
MRP1 inhibitor MK571 or the GSH-depleting agent
buthionine sulphoximine, both cell lines became more sensitive and showed decreased survival. Transport experiments were performed with the (3)H-labelled
glutathione S-conjugate of 4HNE ([(3)H]GS-4HNE) with membrane vesicles from GLC(4)-derived cell lines with different expression levels of
MRP1. [(3)H]GS-4HNE was taken up in an
ATP-dependent manner and the transport rate was dependent on the amount of
MRP1. The
MRP1 inhibitor MK571 decreased [(3)H]GS-4HNE uptake. MRP1-specific [(3)H]GS-4HNE transport was demonstrated with membrane vesicles from High Five insect cells overexpressing recombinant
MRP1. Kinetic experiments showed an apparent K(m) of 1.6+/-0.21 microM (mean+/-S.D.) for MRP1-mediated [(3)H]GS-4HNE transport. In conclusion,
MRP1 has a role in the protection against 4HNE toxicity and GS-4HNE is a novel
MRP1 substrate.
MRP1, together with GSH, is hypothesized to have a role in the defence against oxidative stress.