GENE REGULATION IN PHORBOL SENSITIVE AND RESISTANT MICE

佛波醇敏感和耐药小鼠的基因调控

• 批准号: 3197206
• 负责人: ANDREW P BUTLER
• 金额: $ 18.99万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-07 至 1995-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3197206
• 关键词: casein kinase; chemical carcinogenesis; cytokine; drug resistance; enzyme induction /repression; enzyme mechanism; genetic transcription; genetic translation; growth factor; isozymes; laboratory mouse; neoplasm /cancer genetics; oncogenes; ornithine decarboxylase; phorbols; posttranslational modifications; protein kinase C; protooncogene; skin neoplasms; tissue /cell culture; tumor promoters; ultracentrifugation

The mouse skin multistage carcinogenesis model provides a useful system for the identification and characterization of the sequential events which occur during carcinogenesis. We have found that the epidermis from mice "sensitive" or "resistant" to phorbol ester tumor promotion (SSIN and C57BL/61, respectively) each displayed a large induction of ornithine decarboxylase (ODC) when treated with a single dose of 12-O- tetradecanoylphorbol-13-acetate (TPA), while a second TPA treatment 24 hr later did not result in ODC induction (refractory period; 8). Protein kinase C (PKC) was down-regulated at these times. However, by 48 hr (C57B1/6J) or 72 hr (SSIN), a responsive state had returned in which ODC was over-expressed, even though PKC activity levels continued to decrease. The goal of the proposed research is to elucidate the mechanisms by which the induction of ODC and of nuclear proto-oncogenes (c-jun and c-fos) is dissociated from the activation of PKC during multi-stage carcinogenesis in the mouse epidermis. Towards this aim, we will analyze the signal transduction pathways leading to over-expression of ODC in C57BL/6J and SSIN mice during periods of PKC depletion following a single application of TPA to the epidermis, and will also compare the process of ODC induction by PKC activators (TPA) and promoting agents which induce ODC by non-PKC mediated mechanisms (ethyl phenylpropiolate, EPP). These objectives will be pursued through the following specific aims: (1) Determine the status of protein kinase C activity and isozymes, as well as phorbol ester receptors in the epidermis of tumor promotion sensitive and resistant mice at times following TPA treatment when ODC is over-expressed by a subsequent TPA or EPP treatment. (2) Investigate the roles of nuclear oncogenes c- fos and c-jun in the over induction of ODC by TPA and EPP during periods of PKC depletion. (3) Determine whether the over-expression of ODC observed following a second treatment of skin with TPA is due to augmented transcription, translation, or post-translational effects, using Northern and Western blot techniques and ODC enzyme kinetic determinations. (4) Examine the potential role of casein kinase II and S6 kinase as intermediates in the over-expression of ODC during PKC depletion in the skin. (5) Determine the possible involvement of growth factors or cytokines in the over-induction of ODC while PKC is depleted, using an in vitro cell culture model.

小鼠皮肤多阶段癌变模型为 顺序事件的识别和表征 发生在致癌过程中。 我们发现来自小鼠的表皮 对凤凰酯肿瘤促进的“敏感”或“抗性”（SSIN和SSIN和 C57BL/61分别显示出鸟嘌呤的大诱导 脱羧酶（ODC）用单剂量12-O-处理 tetradecanoylphorbol-13-乙酸酯（TPA），而第二次TPA处理24小时 后来没有导致ODC诱导（难治性周期； 8）。 蛋白质 激酶C（PKC）在这些时候下调。 但是，到48小时 （C57B1/6J）或72小时（SSIN），响应迅速的状态已返回该ODC 即使PKC活性水平继续降低，也表现得很过表达。 拟议研究的目的是阐明 ODC和核原始基因（C-Jun和C-Fos）的诱导是 在多阶段致癌过程中与PKC的激活分离 小鼠表皮。 为了实现这一目标，我们将分析信号 导致C57BL/6J中ODC过度表达的转导途径，并且 单一应用后，PKC耗竭期间的SSIN小鼠 TPA与表皮，还将比较通过 PKC激活剂（TPA）和促进剂，可诱导非PKC ODC 介导的机制（苯基丙酸乙酯，EPP）。 这些目标将 通过以下特定目的追求：（1）确定状态 蛋白激酶C活性和同工酶以及佛波酯的 肿瘤促进敏感和抗性小鼠表皮中的受体 在TPA处理后，有时会被ODC过表达。 TPA或EPP治疗。 （2）研究核癌基因C-的作用 在TPA和EPP过度诱导ODC中的FOS和C-JUN期间 PKC耗竭。 （3）确定是否观察到ODC的过表达 在第二次使用TPA治疗皮肤之后，是由于增强 使用北部的转录，翻译或翻译后效应 以及蛋白质印迹技术和ODC酶动力学测定。 （4） 研究酪蛋白激酶II和S6激酶的潜在作用 PKC耗竭期间ODC过表达的中间体 皮肤。 （5）确定生长因子或 PKC耗尽的ODC过度诱导中的细胞因子，使用IN 体外细胞培养模型。