权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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>基因在皮肤癌变过程中的作用，我们试图通过生物发光技术建立小鼠模型，观察p16^<INK4a>基因在活体小鼠中的表达。我们利用含有约200kbp基因组DNA的BAC克隆，包括p16^<INK4a>基因位点，并在p16^<INK4a>基因最后一个编码外显子的下游插入荧光素酶基因，构建了重组BAC克隆。利用这个重组BAC克隆，我们已经产生了一个转基因小鼠系，产生p16-荧光素酶融合蛋白，以观察p16^<INK4a>在活小鼠中的表达。为了观察p16^<INK4a> -荧光素酶融合的表达，我们给小鼠注射荧光素（荧光素酶的底物），麻醉下在CCD相机下暴露15分钟。我们证实了来自各个器官的光发射与内源性p16^<INK4a>基因的表达有很好的相关性，p16^<INK4a>基因是最重要的肿瘤抑制基因之一，在超过50%的人类癌症中是不活跃的。p16^<INK4a>基因可被激活的Ras信号等致癌信号诱导，是抗肿瘤的失效安全机制。在本研究中，我们研究了p16^<INK4a>基因在体内如何参与和诱导皮肤癌的发生。利用上述转基因小鼠，对p16^<INK4a>基因进行实时可视化。我们使用化学诱导的皮肤癌变模型，采用dba - tpa方案，在小鼠皮肤中形成皮肤乳头状瘤。已知DMBA可诱导H-Ras基因突变，激活Ras信号。小鼠的生物发光在晚期乳头瘤（dba - tpa治疗开始后12周）中增加。我们证实了生物发光的强度与内源性p16^<INK4a>基因在乳头瘤中的表达有良好的相关性，这些结果表明p16^<INK4a>基因在晚期乳头瘤中是被诱导的，并且它抑制肿瘤从良性到恶性的进展较少