Relationship between 4-hydroxyanisole toxicity and dopa oxidase activity for three melanoma cell lines.
We studied the response of mouse B16F10 and SK-MEL-28 and SK-MEL-1 human melanoma cell lines to treatment with 4-hydroxyanisole (4-HA), and attempted to relate the response to the dopa oxidase levels and the morphological characteristics of each cell line. Clear dose-response curves were observed after 24 h of treatment in each cell line, the 4-HA being more toxic to the B16F10 cells, with an ID50 value of 215 microM. This was much lower than that observed for the SK-MEL-28 and SK-MEL-1 cell lines (ID50 of 5.98 mM and 7.17 mM, respectively). There was a direct relationship between toxicity levels and dopa oxidase activity, since the highest specific activity was obtained for B16F10 (15.9 mU), while lower activity was registered for SK-MEL-28 (4.59 mU) and SK-MEL-1 (1.24 mU), which also showed lower 4-HA toxicity. Morphologically, we observed the typical characteristics of cellular injury, with swelling and dilation of the internal membranes and organelles, an increased number of vacuoles, and an increased number of abnormal multilamellar melanosomes or thick clumps of irregularly distributed melanin. On the other hand, we observed that the two cell lines with the lowest dopa oxidase activity contained more mature fully melanized melanosomes than B16F10, pointing to possible alterations in the melanosome transference mechanism and lower enzymatic activity in the mature melanosomes of these two human cell lines.
Rodriguez-Vicente
J
J
Department of Pathology, Faculty of Medicine, University of Murcia, Spain. jrv@fcu.um.es
Vicente-Ortega
V
V
Canteras-Jordana
M
M
Calderon-Rubiales
F
F
D000818:Animals; D000840:Anisoles / Q000633:toxicity*; D000970:Antineoplastic Agents / Q000633:toxicity*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007700:Kinetics; D008545:Melanoma / Q000201:enzymology* / Q000473:pathology / Q000648:ultrastructure; D008546:Melanoma, Experimental / Q000201:enzymology* / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D015388:Organelles / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure; D012878:Skin Neoplasms / Q000201:enzymology* / Q000473:pathology / Q000648:ultrastructure; D014407:Tumor Cells, Cultured
D000840:Anisoles; D000970:Antineoplastic Agents; C009760:mequinol; D014442:Monophenol Monooxygenase