cDNA-array profiling of melanomas and paired melanocyte cultures

J Cell Physiol. 2006 Jun;207(3):697-705. doi: 10.1002/jcp.20610.

Abstract

Three paired (from the same donor) sets of melanoma cells and normal melanocytes, established as early-passage cultures from metastatic lesions and the uninvolved skin of three patients, were comparatively cDNA profiled by macroarrays (approximately 1,200 genes) and reverse transcription (RT)-PCR. While 145 gene products were significantly (at least twofold) upregulated or downregulated in at least 1 pair, and 23 were in at least 2 pairs, only 3 (the signal transducer and activator of transcription STAT2, collagen type VI, and CD9) were concordantly modulated (downregulation) in all 3 pairs. Array results were validated by RT-PCR on a small panel of surgically removed nevocellular nevi and metastatic melanoma lesions, and by immunohistochemistry on a large panel of benign and malignant lesions of the nevomelanocytic lineage. The three gene products were downregulated at different stages of melanoma progression. STAT2 was detectable in nevi (5/5) and most primary melanomas (11/12), but was lost in 10/15 metastatic lesions. Collagen type VI was expressed in nevi (5/5) and primary melanomas below a Breslow thickness of 1 mm (3/3), but was lost in 24/24 primary melanomas above this threshold, and in metastatic melanomas (10/10). The tetraspanin CD9 molecule was expressed in 18/18 nevi, but was lost in 20/28 primary melanomas (including thin lesions), and in 24/52 metastatic lesions. These data provide the proof of principle that cDNA profiling of paired melanocyte/melanoma cultures sorts out novel, early signatures of melanocyte transformation that could contribute to the clinical management of patients at high risk of metastatic disease.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / metabolism
  • Cell Lineage
  • Cells, Cultured
  • Collagen Type IV / metabolism
  • Down-Regulation / genetics
  • Gene Expression Profiling*
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Immunohistochemistry
  • Melanocytes / cytology*
  • Melanocytes / metabolism*
  • Melanoma / genetics*
  • Melanoma / pathology*
  • Membrane Glycoproteins / metabolism
  • Neoplasm Staging
  • Oligonucleotide Array Sequence Analysis*
  • RNA, Messenger / genetics
  • STAT2 Transcription Factor / metabolism
  • Tetraspanin 29
  • Up-Regulation / genetics

Substances

  • Antigens, CD
  • CD9 protein, human
  • Collagen Type IV
  • Membrane Glycoproteins
  • RNA, Messenger
  • STAT2 Transcription Factor
  • STAT2 protein, human
  • Tetraspanin 29