Schlingemann Joerg, Hess Jochen, Wrobel Gunnar, Breitenbach Ute, Gebhardt Christoffer, Steinlein Peter, Kramer Heidi, Fürstenberger Gerhard, Hahn Meinhard, Angel Peter, Lichter Peter
Division of Molecular Genetics, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Int J Cancer. 2003 May 10;104(6):699-708. doi: 10.1002/ijc.11008.
Malignant transformation of mouse skin by chemical carcinogens and tumour promoters, such as the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), is a multistage process that leads to squamous cell carcinoma (SCC) formation. In an effort to identify tumour-associated genes, we studied the influence of short-term TPA-treatment on the gene expression profile of murine skin. A comprehensive microarray with some 5,000 murine gene specific cDNA fragments was established and hybridised with pooled RNA derived from control and TPA-treated dorsal skin samples. Of these genes, 54 were up- and 35 were down-regulated upon TPA application. Additionally, we performed suppression subtractive hybridisation (SSH) with respective RNA pools to generate and analyse a cDNA library enriched for TPA-inducible genes. Expression data of selected genes were confirmed by quantitative real-time PCR and Northern blot analysis. Comparison of microarray and SSH data revealed that 26% of up-regulated genes identified by expression profiling matched with those present in the SSH library. Besides numerous known genes, we identified a large set of unknown cDNAs that represent previously unrecognised TPA-regulated genes in murine skin with potential function in tumour promotion. Additionally, some TPA-induced genes, such as Sprr1A, Saa3, JunB, Il4ralpha, Gp38, RalGDS and Slpi exhibit high basal level in advanced stages of skin carcinogenesis, suggesting that at least a subgroup of the identified TPA-regulated genes may contribute to tumour progression and metastasis.
化学致癌物和肿瘤促进剂（如佛波酯12 - O - 十四酰佛波醇 - 13 - 乙酸酯（TPA））诱导小鼠皮肤发生恶性转化是一个多阶段过程，最终导致鳞状细胞癌（SCC）形成。为了鉴定肿瘤相关基因，我们研究了短期TPA处理对小鼠皮肤基因表达谱的影响。构建了一个包含约5000个小鼠基因特异性cDNA片段的综合微阵列，并与来自对照和TPA处理的背部皮肤样本的混合RNA进行杂交。在这些基因中，TPA处理后有54个基因上调，35个基因下调。此外，我们用相应的RNA池进行抑制性消减杂交（SSH），以生成和分析一个富含TPA诱导基因的cDNA文库。通过定量实时PCR和Northern印迹分析对所选基因的表达数据进行了确认。微阵列和SSH数据的比较显示，通过表达谱分析鉴定出的上调基因中有26%与SSH文库中的基因匹配。除了众多已知基因外，我们还鉴定出大量未知的cDNA，它们代表了小鼠皮肤中以前未被识别的TPA调节基因，在肿瘤促进中可能具有潜在功能。此外，一些TPA诱导的基因，如Sprr1A、Saa3、JunB、Il4ralpha、Gp38、RalGDS和Slpi在皮肤癌发生的晚期表现出高基础水平，这表明至少所鉴定的TPA调节基因的一个亚组可能有助于肿瘤进展和转移。
