Mischiati Carlo, Natali Pier Giorgio, Sereni Alessia, Sibilio Leonardo, Giorda Ezio, Cappellacci Sandra, Nicotra Maria Rita, Mariani Giustino, Di Filippo Franco, Catricalà Caterina, Gambari Roberto, Grammatico Paola, Giacomini Patrizio
Department of Biochemistry and Molecular Biology, University of Ferrara, Italy.
J Cell Physiol. 2006 Jun;207(3):697-705. doi: 10.1002/jcp.20610.
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.
从三名患者的转移病灶和未受累皮肤中建立的三对（来自同一供体）黑色素瘤细胞和正常黑素细胞，作为早期传代培养物，通过宏阵列（约1200个基因）和逆转录（RT）-PCR进行比较cDNA分析。虽然145种基因产物在至少1对中显著（至少两倍）上调或下调，23种在至少2对中显著上调或下调，但只有3种（信号转导和转录激活因子STAT2、VI型胶原和CD9）在所有3对中均被一致调节（下调）。通过对一小部分手术切除的痣细胞痣和转移性黑色素瘤病灶进行RT-PCR，以及对一大组痣细胞系的良性和恶性病灶进行免疫组织化学，验证了阵列结果。这三种基因产物在黑色素瘤进展的不同阶段下调。STAT2在痣（5/5）和大多数原发性黑色素瘤（11/12）中可检测到，但在10/15的转移病灶中消失。VI型胶原在痣（5/5）和Breslow厚度低于1mm的原发性黑色素瘤（3/3）中表达，但在超过此阈值的24/24原发性黑色素瘤以及转移性黑色素瘤（10/10）中消失。四跨膜蛋白CD9分子在18/18的痣中表达，但在20/28的原发性黑色素瘤（包括薄病灶）和24/52的转移病灶中消失。这些数据提供了原理证明，即配对黑素细胞/黑色素瘤培养物的cDNA分析筛选出了黑素细胞转化的新的早期特征，这可能有助于对有转移性疾病高风险患者的临床管理。
