ENERGY RESTRICTION, CELL SIGNALING AND CANCER PREVENTION

描述：拟议的研究将测试饮食 能量限制（ER）通过提高 糖皮质激素（GCH），其减少选定的表皮蛋白激酶 C（PKC α和ζ）并阻断AP-1诱导。 第一个具体目标 将确定肾上腺是否需要ER抑制肿瘤 如果对肾上腺切除/ER小鼠施用GCH， ER抑制。 皮肤肿瘤将开始与7，12-二甲基 苯并（a）蒽（DMBA），并用TPA促进。 GCH的高度为 假设其通过引起皮肤肿瘤促进来介导抑制皮肤肿瘤促进 PKC蛋白水平的降低抑制了 Raf-1/MAP-激酶通路并阻断TPA诱导c-June的能力 蛋白质和AP-1：DNA结合。 具体目标2将检验以下假设： AP-1：DNA结合诱导的阻断依赖于ER AP-1组成蛋白的调节。 第三个具体目标是 检验ER阻断Raf-1/MAP激酶信号传导的假设 Raf-1是PKC的一种已知底物。 凝胶迁移率 使用AP-1 DNA共有序列的移位实验、蛋白质印迹和 免疫复合物激酶测定将用于目的2和3。 第四 特定目标，饮食能量限制阻断AP-1的能力 荧光素酶在TPA和UVB光下诱导和转录调控 将使用转基因AP-1-荧光素酶小鼠研究治疗的小鼠 模型 目的2-4中修改的生化事件对以下的依赖性： 皮质酮将通过在假手术和 肾上腺切除小鼠。

DESCRIPTION: The proposed studies will test the hypothesis that dietary energy restriction (ER) inhibits skin tumor promotion by elevating glucocorticoid hormone (GCH) which reduces selected epidermal protein kinase C (PKC alpha and zeta) and blocks AP-1 induction. The first specific aim will determine if the adrenal gland is required for ER inhibition of tumor promotion and if administration of GCH to adrenalectomized/ER mice restores the ER inhibition. Skin tumors will be initiated with 7,12-dimethyl benz(a)anthracene (DMBA) and promoted with TPA. Elevation of GCH is hypothesized to mediate the inhibition of skin tumor promotion by causing the reduction in PKC protein levels which inhibits signaling down Raf-1/MAP-Kinase pathways and blocks the ability of TPA to induce c-June protein and AP-1:DNA binding. Specific Aim 2 will test the hypothesis that the blockage of the induction of AP-1:DNA binding is dependent upon ER regulation of constituent proteins of AP-1. The third specific aim will test the hypothesis that ER blocks signaling down the Raf-1/MAP-kinase pathway at the level of Raf-1, a well known substrate for PKC. Gel mobility shift experiments with AP-1 DNA consensus sequences, western blots and immune complex kinase assays will be used for Aims 2 and 3. In the fourth specific aim, the ability of dietary energy restriction to block AP-1 induction and transcriptional regulation of luciferase in TPA and UVB-light treated mice will be investigated using the transgenic AP-1-luciferase mouse model. The dependence of the biochemical events modified in Aims 2-4 on corticosterone will be assessed by parallel studies in sham and adrenalectomized mice.