Glutathione protects metastatic melanoma cells against oxidative stress in the murine hepatic microvasculature.
Calcein-labeled B16 melanoma (B16M) cells were injected intraportally, and in vivo video microscopy was used to study the distribution and damage of cancer cells arrested in the liver microvasculature over a period of 4 hours. The contribution of glutathione (GSH)-dependent antioxidant machinery to the possible oxidative stress-resistance mechanism of B16M cell was determined by in vitro incubation with the selective inhibitor of GSH synthesis L-buthionine (S,R)-sulphoximine (BSO) before B16M cell injection in untreated and 0.5-mg/kg lipopolysaccharide (LPS)-treated mice. In addition, untreated and LPS-treated isolated syngeneic hepatic sinusoidal endothelial cells (HSE) were used to determine in vitro their specific contribution to B16M cell damage. Trauma inherent to intrasinusoidal lodgement damaged 35% of B16M cells in both normal and LPS-treated mouse liver. The rest of the arrested B16M cells remained intact in normal liver for at least 4 hours, although their damaged cell percentage significantly (P < .05) increased since the second hour in normal mice injected with BSO-treated cells and since the first hour in LPS-treated mice given untreated cells. Recombinant human interleukin-1 receptor antagonist (rHuIL-1-Ra) given to mice 15 minutes before LPS significantly (P < .05) abrogated B16M cell damage. On the other hand, 40% of the B16M cells co-cultured with unstimulated HSE and 70% of the co-cultured with LPS-treated HSE became sensitive to endothelial cell-mediated damage after BSO treatment. These results demonstrate that a high intracellular level of GSH protects B16M cells from possible in vivo and in vitro sinusoidal cell-mediated oxidative stress, contributing to the mechanism of metastatic cell survival within the hepatic microvasculature.
Anasagasti
M J
MJ
Department of Cell Biology and Morphological Sciences, University of the Basque Country, School of Medicine and Dentistry, Leioa, Vizcaya, Spain.
Martin
J J
JJ
Mendoza
L
L
Obrador
E
E
Estrela
J M
JM
McCuskey
R S
RS
Vidal-Vanaclocha
F
F
D000818:Animals; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002448:Cell Adhesion; D016764:Cell Polarity; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005978:Glutathione / Q000502:physiology*; D008099:Liver / Q000098:blood supply / Q000473:pathology; D008113:Liver Neoplasms / Q000556:secondary; D008297:Male; D008546:Melanoma, Experimental / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008833:Microcirculation; D009362:Neoplasm Metastasis; D018384:Oxidative Stress / Q000187:drug effects*; D014407:Tumor Cells, Cultured; D014741:Video Recording
D004791:Enzyme Inhibitors; D019328:Buthionine Sulfoximine; D005978:Glutathione