title: Identification of differentially expressed genes in uveal melanoma using suppressive subtractive hybridization.
abstract: OBJECTIVE
  Uveal melanoma (UM) is the most common primary cancer of the eye, resulting not only in vision loss, but also in metastatic death. This study attempts to identify changes in the patterns of gene expression that lead to malignant transformation and proliferation of normal uveal melanocytes (UVM) using the Suppressive Subtractive Hybridization (SSH) technique.
  
  
  METHODS
  The SSH technique was used to isolate genes that are differentially expressed in the TP31 cell line derived from a primary UM compared to UVM. The expression level of selected genes was further validated by microarray, semi-quantitative RT-PCR and western blot analyses.
  
  
  RESULTS
  Analysis of the subtracted libraries revealed that 37 and 36 genes were, respectively, up- and downregulated in TP31 cells compared to UVM. Differential expression of the majority of these genes was confirmed by comparing UM cells with UVM by microarray. The expression pattern of selected genes was analyzed by semi-quantitative RT-PCR and western blot, and was found to be consistent with the SSH findings.
  
  
  CONCLUSIONS
  We demonstrated that the SSH technique is efficient to detect differentially expressed genes in UM. The genes identified in this study represent valuable candidates for further functional analysis in UM and should be informative in studying the biology of this tumor.
authors.affiliation: LOEX/CUO Recherche, Centre Hospitalier Affilié Universitaire de Québec, Hôpital du Saint-Sacrement, and Département d'Ophtalmologie, Faculté de Médecine, Université Laval, Québec, Québec, Canada.
authors.forename: Solange
authors.initials: S
authors.lastname: Landreville
authors.forename: Caroline B
authors.initials: CB
authors.lastname: Lupien
authors.forename: Francois
authors.initials: F
authors.lastname: Vigneault
authors.forename: Manon
authors.initials: M
authors.lastname: Gaudreault
authors.forename: Mélissa
authors.initials: M
authors.lastname: Mathieu
authors.forename: Alain P
authors.initials: AP
authors.lastname: Rousseau
authors.forename: Sylvain L
authors.initials: SL
authors.lastname: Guérin
authors.forename: Christian
authors.initials: C
authors.lastname: Salesse
mesh_terms: D000328:Adult; D000368:Aged; D015153:Blotting, Western; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D055028:Comparative Genomic Hybridization / Q000379:methods*; D015536:Down-Regulation; D005123:Eye / Q000378:metabolism / Q000473:pathology*; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D052138:Genes, Neoplasm; D006801:Humans; D008544:Melanocytes / Q000378:metabolism*; D008545:Melanoma / Q000235:genetics / Q000473:pathology; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation; D014604:Uveal Neoplasms / Q000235:genetics / Q000473:pathology; D000098943:Uveal Melanoma
chemical_list: D012333:RNA, Messenger