Hypoxia-inducible factor1a in multistage carcinogenesis

描述（申请人提供）：缺氧诱导因子1-α（HIF-1 α） 是转录因子HIF-1的组成部分。细胞培养研究和 小鼠基因敲除证实HIF-1 α反式激活基因控制 血管生成、细胞代谢、侵袭和凋亡。除了 缺氧HIF-1 α蛋白表达受生长因子信号传导调节， 生存途径和病毒致癌基因。HIF-1 α在癌症中被诱导表达 和高级别癌前病变然而. HIF-1 α在乳腺癌各阶段的作用 上皮癌发生或转移是未知的。我们创造了转基因小鼠 表达野生型或缺乏组成型活性突变HIF-1 α， 基底鳞状上皮中存在“氧依赖性降解结构域 （K14-HIF-1 α和K14-HIF-1 α δ ODD转基因小鼠）。我们设计了 Cre-loxP介导的基底鳞状上皮中HIF-1a缺失， 可辨别的表型现在，我们测试HIF-1 α是否在 具有这些特定目的的多阶段上皮癌发生 1.确定HIF-1 α功能的获得或丧失对 Ha-ras启动了表皮癌的发生。1.1.检测HIF-1 α的增加是否 功能促进两阶段表皮化学致癌诱导 二甲基苯并蒽和肉豆蔻酸四佛波酯乙酸酯， Kl 4-HIF-laz\ODD转基因小鼠。1.2. HIF-1 α两阶段检测的必要性 Cre-loxP介导的表皮HIF-1 α对小鼠的化学致癌作用 删除。2.0.确定多阶段HIF-1 α功能获得的生物学 人乳头瘤病毒16型早期转化诱发皮肤和宫颈癌的研究 地区2.1. HPV 16诱导的表皮细胞各阶段的试验改变 KI 4-HPV 16中HIF-1 α功能获得致癌作用：HIF-1 α或HIF-1 α δ ODD双转基因小鼠。2.2.测试HIF-1 α之间的合作增益 雌激素在HPV 16诱发小鼠宫颈癌中的作用 HPV 16和HIF-1 α或HIF-1 α δ ODD转基因。3.0.试验生物学 HPV 16 E6-HIF-1 α共表达，并确定p53依赖或不依赖 E6与HIF-1 α的功能。3.1.创建Kl 4-E6 HIF-1 α或 K14-E6：H1 F-1 α δ ODD双转基因小鼠，并测定H1 F-1 α δ ODD双转基因小鼠中的改变。 皮肤和宫颈致癌生物学。3.2.创建Kl 4-E6 HIF-1 α或 K14-E6：HIF-1 α δ ODD双转基因小鼠，并确定其基因表达的改变。 皮肤和宫颈致癌生物学。我们拥有所有相关的 动物模型，使我们能够确定HIF-1 α在 由激活的癌基因或肿瘤失活介导的致癌作用 抑制基因

DESCRIPTION (PROVIDED BY APPLICANT):Hypoxia inducible factor 1-alpha (HIF-1alpha) is a component of the transcription factor HIF-1. Cell culture studies and mouse gene knockouts demonstrate HIF-1alpha transactivation of genes controlling angiogenesis, cellular metabolism, invasion and apoptosis. In addition to hypoxia. HIF-1alpha protein expression is regulated by growth factor signaling and survival pathways, and viral oncogenes. HIF-1alpha expression is induced in cancers and high-grade premalignant lesions. However. HIF-1alpha function in each stage of epithelial carcinogenesis or metastasis is unknown. We created transgenic mice expressing either wild type or a constitutively active mutant HIF-1alpha lacking the "oxygen-dependent degradation domain' about in basal squamous epithelium (K14-HIF-1alpha and K14-HIF-1alpha delta ODD transgenic mice). We have engineered Cre-loxP mediated HIF-1a deletion in basal squamous epithelium without a discernable phenotype. Now we test whether HIF-1alpha plays a fundamental role in multistage epithelial carcinogenesis with these Specific Aims 1. Determine the stage-specific effect of gain or loss of HIF-1alpha function on Ha-ras initiated epidermal carcinogenesis. 1.1. Test whether gain of HIF- 1alpha function promotes two-stage epidermal chemical carcinogenesis induced by dimethylbenzanthracene and tetraphorbolmyristate acetate in Kl4-HIF-1alpha and Kl4-HIF-laz\ODD transgenic mice. 1.2. Test necessity of HIF-1alpha in two-stage chemical carcinogenesis in mice with Cre-loxP mediated epidermal HIF-1alpha deletion. 2.0. Determine biology of gain of HIF-1alpha function on multistage carcinogenesis in skin and cervix induced by the HPV16 early transforming region. 2.1. Test alteration of each stage of HPV16-induced epidermal carcinogenesis by gain of HIF-1alpha function in KI4-HPV16:HIF-1alpha or HIF-1alpha delta ODD double transgenic mice. 2.2. Test cooperation between HIF-1alpha gain of function and estrogen in HPV16 induced cervical carcinogenesis in mice transgenic for the HPV16 and HIF-1alpha or HIF-1alpha delta ODD. 3.0. Test biology of HPV16 E6-HIF-1alpha coexpression, and determine p53 dependent or independent functions of E6 in conjunction with HIF-1alpha. 3.1. Create Kl4-E6HIF-1alpha or K14-E6:HlF-1alpha delta ODD double transgenic mice and determine alterations in the biology of skin and cervical carcinogenesis. 3.2. Create Kl4-E6 HIF-1alpha or K14-E6:HIF-1alpha delta ODD double transgenic mice and determine alterations in the biology of skin and cervical carcinogenesis. Our possession of all relevant animal models, positions us to determine the precise role of HIF-1alpha in carcinogenesis mediated by either activated oncogenes or inactivation of tumor suppressor genes.