Eberle J, Garbe C, Orfanos C E
Freie Universität Berlin, Klinikum Steglitz Hautklinik und Poliklinik, Germany.
Arch Dermatol Res. 1995;287(5):421-7. doi: 10.1007/BF00373422.
In order to improve the characterization of human malignant melanoma cells and their variant gene expression in vitro, a search for specifically regulated genes was performed. Four melanoma cell lines (M5, MEWO, IGR39, SKMEL13) and newly cultured normal human melanocytes were included in a comparative hybridization (differential screening) of a human melanoma cDNA-library. Six cDNAs were isolated showing a stronger expression (four genes) or a weaker expression (two genes) in melanoma cells than in normal human melanocytes. Quantification of the expression patterns of the two repressed genes in Northern blots revealed general expression in all melanocyte cultures examined, no expression in three cell lines (M5, IGR39, SKMEL13) and weak expression in MEWO. The four induced genes were found to be only weakly expressed in normal human melanocytes, but showed an elevated expression in all of the four melanoma cell lines tested. Thus, using the technique of differential screening, consistent gene regulation at the messenger RNA level was identified, which distinguishes the four melanoma cell lines tested from normal melanocytes. We conclude from the expression patterns that specific gene regulation in melanoma cells in vitro is characterized both by strong repression of some melanocyte genes, as well as by the induction of other genes, but there was no indication of new expression of genes specific for melanoma cells. Because of the uniform induction or repression in different melanoma cell lines, it is conceivable that the cloned genes may be involved in the malignant transformation of melanocytic cells.
为了在体外更好地表征人类恶性黑色素瘤细胞及其变异的基因表达，我们开展了对特异性调控基因的搜索。将四种黑色素瘤细胞系（M5、MEWO、IGR39、SKMEL13）和新培养的正常人黑色素细胞纳入人黑色素瘤cDNA文库的比较杂交（差异筛选）。分离出六个cDNA，它们在黑色素瘤细胞中的表达比在正常人黑色素细胞中更强（四个基因）或更弱（两个基因）。对Northern印迹中两个受抑制基因的表达模式进行定量分析，结果显示在所检测的所有黑色素细胞培养物中均有表达，在三个细胞系（M5、IGR39、SKMEL13）中无表达，在MEWO中表达较弱。发现四个诱导基因在正常人黑色素细胞中仅微弱表达，但在所有四个测试的黑色素瘤细胞系中表达升高。因此，通过差异筛选技术，在信使RNA水平鉴定出了一致的基因调控，这将所测试的四个黑色素瘤细胞系与正常黑色素细胞区分开来。从表达模式我们得出结论，体外黑色素瘤细胞中的特异性基因调控既表现为一些黑色素细胞基因的强烈抑制，也表现为其他基因的诱导，但没有迹象表明存在黑色素瘤细胞特异性新基因的表达。由于在不同黑色素瘤细胞系中诱导或抑制情况一致，可以推测克隆的基因可能参与黑色素细胞的恶性转化。
