Wei Sung-Jen, Trempus Carol S, Cannon Ronald E, Bortner Carl D, Tennant Raymond W
National Center for Toxicogenomics and the Laboratory of Signal Transduction, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709, USA.
J Biol Chem. 2003 Jan 17;278(3):1758-68. doi: 10.1074/jbc.M206328200. Epub 2002 Nov 4.
This study identifies genes expressed early in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced skin carcinogenesis in genetically initiated Tg.AC v-Ha-ras transgenic mice. Keratinocyte progenitor cells from TPA-treated Tg.AC mice were isolated with fluorescence-activated cell sorting and expression was analyzed using cDNA microarray technology. Eleven genes were identified whose expression changed significantly in response to carcinogen treatment. Deleted in split hand/split foot 1 (Dss1) is a gene associated with a heterogeneous limb developmental disorder called split hand/split foot malformation. cDNA microarray expression analysis showed that the mouse homologue of Dss1 is induced by TPA. Dss1 overexpression was detected by Northern blot analysis in early TPA-treated hyperplastic skins and in JB6 Cl 41-5a epidermal cells. Interestingly, Dss1 expression was also shown to be elevated in skin papillomas relative to normal skins, and further increased in squamous cell malignancies. Functional studies by ectopically constitutive expression of Dss1 in JB6 Cl 41-5a preneoplastic cells strongly increased focus formation and proliferation of these cells and enhanced efficiency of neoplastic transformation of the cells in soft agar. These results strongly suggest that Dss1 is a TPA-inducible gene that may play an important role in the early stages of skin carcinogenesis.
本研究鉴定了在遗传启动的Tg.AC v-Ha-ras转基因小鼠中,12-氧十四烷酰佛波醇-13-乙酸酯(TPA)诱导皮肤癌发生早期表达的基因。用荧光激活细胞分选法从经TPA处理的Tg.AC小鼠中分离角质形成细胞祖细胞,并使用cDNA微阵列技术分析其表达。鉴定出11个基因,其表达在致癌物处理后发生显著变化。手足裂畸形1缺失基因(Dss1)是一种与名为手足裂畸形的异质性肢体发育障碍相关的基因。cDNA微阵列表达分析表明,Dss1的小鼠同源物由TPA诱导。通过Northern印迹分析在早期经TPA处理的增生性皮肤和JB6 Cl 41-5a表皮细胞中检测到Dss1过表达。有趣的是,相对于正常皮肤,皮肤乳头瘤中Dss1表达也升高,在鳞状细胞恶性肿瘤中进一步增加。通过在JB6 Cl 41-5a肿瘤前体细胞中异位组成性表达Dss1进行功能研究,强烈增加了这些细胞的集落形成和增殖,并提高了细胞在软琼脂中的肿瘤转化效率。这些结果强烈表明,Dss1是一种TPA诱导基因,可能在皮肤癌发生的早期阶段起重要作用。