Induction of p16^<INK4a> gene expression by oncogenic stress is involved in the tumor suppression mechanism of skin cancer

致癌应激诱导p16^<INK4a>基因表达参与皮肤癌的肿瘤抑制机制

基本信息

批准号：
16390318
负责人：
OHTANI Naoko
金额：
$ 9.22万
依托单位：
University of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390318/
关键词：
p16^<INK4a> gene bioluminescence in vivo imaging skin cancer p16^<Ink4a>遺伝子可視化 BAC

项目摘要

In order to examine the role of p16^<INK4a> gene in the skin carcinogenesis, we tried to generate a mouse model to visualize p 16^<INK4a> gene expression in living mice by bioluminescence. We utilized the BAC clone which contains approximately 200kbp genomic DNA including p16^<INK4a> gene locus, and constructed recombinant BAC clone by inserting luciferase gene in frame at the downstream of the last coding exon of the p16^<INK4a> gene. Using this recombinant BAC clone, we have generated a transgenic mouse line that produces p16-luciferase fusion protein to visualize p16^<INK4a> expression in living mice. To visualize the expression of p16^<INK4a> -Luciferase fusion, mice were subjected with luciferin, the subatrate of luciferase, and exposed for 15 minutes to CCD camera under anesthesia. We confirmed that light emission from each organ is well correlated with the endogenous expression of p16^<INK4a> genep16^<INK4a> gene is one of the most important tumor suppressor genes, and is inacti … More vated in more than 50% of human cancers. p16^<INK4a> gene is known to be induced by oncogenic signal such as activated Ras signal as a fail safe mechanism against oncogenesis. In this study, we investigated how p16^<INK4a> gene is involved and induced in skin cancer development in vivo. By utilizing the transgenic mouse described above, the real-time visualization of p16^<INK4a> gene was performed. We have used the chemical-induced skin carcinogenesis model using DMBA-TPA protocol which develops skin papillomas in the mouse skin. DMBA is known to induce a mutation in H-Ras gene and activate Ras signal. The bioluminescence from the mice was increased in the late papilllomas (later than 12 weeks after DMBA-TPA treatment was started). We confirmed that the intensity of bioluminescence was well correlated with the endogenous p16^<INK4a> gene expression in the papillomas These results suggests that p16^<INK4a> gene is induced in late papillomas, and that it inhibits the tumor progression from benign to malignant skin tumors Less

为了研究 p16^<INK4a> 基因在皮肤癌发生中的作用，我们尝试建立小鼠模型，通过生物发光来可视化活体小鼠中的 p16^<INK4a> 基因表达。我们利用包含约 200kbp 的 BAC 克隆。包括p16^<INK4a>基因位点的基因组DNA，并通过在最后一个编码外显子下游插入荧光素酶基因构建重组BAC克隆。使用该重组 BAC 克隆，我们生成了可产生 p16-荧光素酶融合蛋白的转基因小鼠系，以可视化活体小鼠中 p16^<INK4a> 的表达。 -荧光素酶融合，小鼠接受荧光素酶的底物，并在麻醉下暴露于CCD相机15分钟。每个器官的光发射与p16^<INK4a>基因的内源性表达密切相关p16^<INK4a>基因是最重要的抑癌基因之一，在超过50%的人类癌症中p16基因处于失活状态。已知 ^<INK4a> 基因是由致癌信号（例如激活的 Ras 信号）诱导的，作为对抗肿瘤发生的故障安全机制。在本研究中，我们研究了如何诱导。 p16^<INK4a>基因参与并诱导体内皮肤癌发展通过利用上述转基因小鼠，我们使用化学诱导的皮肤癌发生模型进行了p16^<INK4a>基因的实时可视化。使用 DMBA-TPA 方案在小鼠皮肤中形成皮肤乳头状瘤，已知 DMBA 会诱导 H-Ras 基因突变并激活小鼠的生物发光。乳头状瘤（DMBA-TPA 治疗开始后 12 周后）我们证实生物发光强度与乳头状瘤中的内源性 p16^<INK4a> 基因表达密切相关。这些结果表明 p16^<INK4a> 基因被诱导。在晚期乳头状瘤中，它抑制肿瘤从良性到恶性皮肤肿瘤的进展 Less