title: Influence of melanosome dynamics on melanoma drug sensitivity.
abstract: BACKGROUND
  Malignant melanomas are intrinsically resistant to many conventional treatments, such as radiation and chemotherapy, for reasons that are poorly understood. Here we propose and test a model that explains drug resistance or sensitivity in terms of melanosome dynamics.
  
  
  METHODS
  The growth and sensitivity to cisplatin of MNT-1 cells, which are melanotic and enriched with mature stage III and IV melanosomes, and SK-MEL-28 cells, which have only immature stage I and II melanosomes, were compared using clonogenic assays. Differences in pigmentation, melanosome stages, melanosome number, and cellular structures in different cell lines in response to various treatments were examined by electron microscopy. The relative numbers of melanosomes of different stages were compared after treatment with 1-phenyl-2-thiourea. The relationship between drug transporter function and endogenous melanogenic toxicity was assessed by treating cells with the cyclosporin analog PSC-833 and by assessing vacuole formation and cell growth inhibition. All statistical tests were two-sided.
  
  
  RESULTS
  Endogenous melanogenic cytotoxicity, produced by damaged melanosomes, resulted in pronounced cell growth inhibition in MNT-1 cells compared with amelanotic SK-MEL-28 cells. The sensitivity to CDDP of MNT-1 cells was 3.8-fold higher than that of SK-MEL-28 cells (mean IC(50) for SK-MEL-28 and MNT-1 = 2.13 microM and 0.56 microM, respectively; difference = 1.57 microM, 95% confidence interval = 1.45 to 1.69; P = .0017). After treatment with 6.7 microM CDDP for 72 hours, the number of stage II-III melanosomes in surviving MNT-1 cells was 6.8-fold that of untreated cells. Modulation of MNT-1 cells to earlier-stage (II, II-III, III) melanosomes by treatment with the tyrosinase inhibitor 1-phenyl-2-thiourea dramatically increased CDDP resistance. Furthermore, PSC-833 principally suppressed MNT-1 melanotic cell growth via an elevation of autophagosome-like vacuolar structures, possibly by inhibiting melanosome membrane transporters.
  
  
  CONCLUSIONS
  Melanosome dynamics (including their biogenesis, density, status, and structural integrity) regulate the drug resistance of melanoma cells. Manipulation of melanosome functions may be an effective way to enhance the therapeutic activity of anticancer drugs against melanoma.
authors.affiliation: Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bldg 37, Rm 2108, Bethesda, MD 20892, USA.
authors.forename: Kevin G
authors.initials: KG
authors.lastname: Chen
authors.forename: Richard D
authors.initials: RD
authors.lastname: Leapman
authors.forename: Guofeng
authors.initials: G
authors.lastname: Zhang
authors.forename: Barry
authors.initials: B
authors.lastname: Lai
authors.forename: Julio C
authors.initials: JC
authors.lastname: Valencia
authors.forename: Carol O
authors.initials: CO
authors.lastname: Cardarelli
authors.forename: Wilfred D
authors.initials: WD
authors.lastname: Vieira
authors.forename: Vincent J
authors.initials: VJ
authors.lastname: Hearing
authors.forename: Michael M
authors.initials: MM
authors.lastname: Gottesman
mesh_terms: D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology; D003524:Cyclosporins / Q000494:pharmacology; D004317:Doxorubicin / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D008545:Melanoma / Q000188:drug therapy* / Q000473:pathology*; D008546:Melanoma, Experimental / Q000188:drug therapy / Q000473:pathology; D020460:Melanosomes / Q000187:drug effects* / Q000648:ultrastructure*; D051379:Mice; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D012878:Skin Neoplasms / Q000188:drug therapy* / Q000473:pathology*; D014700:Verapamil / Q000494:pharmacology; D014747:Vinblastine / Q000494:pharmacology
chemical_list: D003524:Cyclosporins; D014747:Vinblastine; D004317:Doxorubicin; D014700:Verapamil; D002945:Cisplatin; C070272:valspodar
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