title: Melanoma, a tumor based on a mutant stem cell?
abstract: Stem cells play a critical role in normal tissue maintenance, and mutations in these stem cells may give rise to cancer. We hypothesize that melanoma develops from a mutated stem cell and therefore residual stem cell characteristics should be able to be identified in melanoma cell lines. We studied three metastatic melanoma cell lines that exhibited multiple morphologic forms in culture and demonstrated the capacity to pigment. We used the ability to efflux Hoechst 33342 dye, a technique known to enrich for stem cells in many tissues, to segregate cell populations. The cells with the greatest ability to efflux the dye were (1) small in size, (2) had the capacity to give rise to larger cell forms, and (3) had the greatest ability to expand in culture. The small cells were found to have a decreased proliferative rate and were less melanized. Large dendritic cells that appeared to be nonproliferative were identified in cultures. Treatment with cytosine beta-D-arabinofuranoside hydrochloride (Ara-C) expanded the large cell population but the residual proliferative capacity, both in vitro and in vivo, remained concentrated in the smaller cell fraction. Antigenic staining patterns were variable and heterogeneous. Nestin (a neural stem cell marker) and gp100 (premelanosomal marker) favored the smaller cell population, while nerve growth factor receptor often labeled larger cells. Morphologic and antigenic heterogeneity remained intact after clonal purification. These findings are consistent with the behavior expected for a tumor based on stem cell biology; this finding has diagnostic and therapeutic implications for melanocytic neoplasias.
authors.affiliation: Department of Medicine, Duke University Medical Center, Durham, North Carolina 27710, USA. grich001@mc.duke.edu
authors.forename: James M
authors.initials: JM
authors.lastname: Grichnik
authors.forename: James A
authors.initials: JA
authors.lastname: Burch
authors.forename: Ryan D
authors.initials: RD
authors.lastname: Schulteis
authors.forename: Siqing
authors.initials: S
authors.lastname: Shan
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.forename: Timothy L
authors.initials: TL
authors.lastname: Darrow
authors.forename: Carol E
authors.initials: CE
authors.lastname: Vervaert
authors.forename: Hilliard F
authors.initials: HF
authors.lastname: Seigler
mesh_terms: D000951:Antigens, Neoplasm / Q000032:analysis; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D001562:Benzimidazoles / Q000032:analysis / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003561:Cytarabine / Q000494:pharmacology; D003713:Dendritic Cells / Q000187:drug effects; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism; D006801:Humans; D007381:Intermediate Filament Proteins / Q000032:analysis; D008545:Melanoma / Q000235:genetics* / Q000473:pathology*; D008562:Membrane Glycoproteins / Q000032:analysis*; D009154:Mutation; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000032:analysis*; D014411:Neoplastic Stem Cells / Q000737:chemistry / Q000473:pathology*; D009419:Nerve Tissue Proteins / Q000032:analysis; D064231:Nestin; D012878:Skin Neoplasms / Q000235:genetics* / Q000473:pathology*; D058951:gp100 Melanoma Antigen
chemical_list: D000951:Antigens, Neoplasm; D000964:Antimetabolites, Antineoplastic; D001562:Benzimidazoles; D005456:Fluorescent Dyes; D007381:Intermediate Filament Proteins; D008562:Membrane Glycoproteins; C577484:NES protein, human; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D064231:Nestin; C548637:PMEL protein, human; D058951:gp100 Melanoma Antigen; D003561:Cytarabine; C017807:bisbenzimide ethoxide trihydrochloride