cDNA-array profiling of melanomas and paired melanocyte cultures.
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.
Mischiati
Carlo
C
Department of Biochemistry and Molecular Biology, University of Ferrara, Italy.
Natali
Pier Giorgio
PG
Sereni
Alessia
A
Sibilio
Leonardo
L
Giorda
Ezio
E
Cappellacci
Sandra
S
Nicotra
Maria Rita
MR
Mariani
Giustino
G
Di Filippo
Franco
F
Catricalà
Caterina
C
Gambari
Roberto
R
Grammatico
Paola
P
Giacomini
Patrizio
P
D015703:Antigens, CD / Q000378:metabolism; D019070:Cell Lineage; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000378:metabolism; D015536:Down-Regulation / Q000235:genetics; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D008544:Melanocytes / Q000166:cytology* / Q000378:metabolism*; D008545:Melanoma / Q000235:genetics* / Q000473:pathology*; D008562:Membrane Glycoproteins / Q000378:metabolism; D009367:Neoplasm Staging; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics; D050795:STAT2 Transcription Factor / Q000378:metabolism; D060245:Tetraspanin 29; D015854:Up-Regulation / Q000235:genetics
D015703:Antigens, CD; C554869:CD9 protein, human; D024141:Collagen Type IV; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D050795:STAT2 Transcription Factor; C494084:STAT2 protein, human; D060245:Tetraspanin 29