PKC ISOZYMES AND SUBSTRATES IN MAMMARY CARCINOGENESIS

描述：细胞和分子功能的变化有助于 乳腺癌的发展和进展在很大程度上是未知的。 生长成分的活性和/或表达水平的变化 与肿瘤相关的调控信号转导通路 已经注意到了进展。了解的功能意义 这些生化关联有望开发出新的和 预防、检测、诊断和治疗的替代战略 乳腺癌。蛋白激酶C(PKC)是一个普遍表达的家族 已知的酶在调节细胞生长的基本机制中起重要作用 和差异化。PKC也被认为是一个重要的信号 肿瘤促进/进展的途径，因为PKC是主要的细胞 肿瘤促进剂佛波酯的受体。这项建议的目的是 是为了清楚地理解PKC同工酶和 正常乳腺细胞转化为多发性硬化症细胞的底物 改变了生长能力，随后进入了一个具有侵袭性的细胞， 转移潜能。PKC实际上是一个不同的同工酶家族， 催化磷脂依赖的蛋白质磷酸化。的作用 细胞过程中的个体同工酶还没有被定义。我们的 初步数据显示，两种PKC同工酶增加，而 与肿瘤转化相关的两种PKC底物 和增加的转移潜能(与D.Medina博士和 D.韦尔奇分别提出)。我们的目标是测试它的功能意义 这些生物化学通过操纵个体的活动而相互关联。 通过表达PKCs、PKC显性负抑制物和 独特的衬底。异位基因将被引入培养细胞中。 表达克隆的肿瘤发病率和转移潜能将 通过将细胞移植到脂肪垫中在体内进行测定。了解以下内容 单个PKC在调节乳腺细胞生长中的作用是 对于开发新的和替代的乳房策略是必不可少的 癌症治疗和预防。具体目标：1.比较 进行性转化的乳腺细胞中的PKC同工酶和底物 肿瘤。2.建立高表达的乳腺细胞系 单个PKC或显性负性抑制因子PKC构建。3.至 确定表达PKCs和PKC显性阴性的效果 抑制乳腺移植细胞肿瘤发生和转移的实验研究 在活体内。4.建立两个PKC高表达的细胞系 底物(克隆34和72)以评估这些蛋白质在生长中的作用 和转移。5.检测表达PKCs、PKC的效果。 对13762只大鼠乳腺细胞的显性负性抑制物和底物 文化。

DESCRIPTION: Changes in cellular and molecular functions that contribute to the development and progression of breast cancer are largely undefined. Changes in activities and/or expression levels of components of growth regulatory signal transduction pathways that correlate with tumor progression have been noted. Understanding the functional significance of these biochemical correlates holds the promise for developing new and alternative strategies for prevention, detection, diagnosis and treatment of breast cancer. Protein kinase C (PKC) is a family of ubiquitously expressed enzymes known to be important in regulating basic mechanisms of cell growth and differentiation. PKC is also considered to be a significant signaling pathway in tumor promotion/progression since PKCs are the major cellular receptors for tumor promoting phorbol esters. The purpose of this proposal is to provide a clear understanding of the role of PKC isozymes and substrates in the transition of a normal breast cell into a cell with altered growth potential and subsequently into a cell with invasive, metastatic potential. PKC is actually a family of distinct isozymes that catalyze phospholipid-dependent protein phosphorylation. The role of individual isozymes in cellular processes has not yet been defined. Our preliminary data demonstrate increases in two PKC isozymes and decreases in two PKC substrates that correlate with conversion of preneoplasias to tumors and increased metastatic potential, (collaborations with Drs. D. Medina and D. Welch, respectively). Our goal is to test the functional significance of these biochemical correlates by manipulating the activities of individual isozymes through expression of PKCs, PKC dominant negative inhibitors and unique substrates. Ectopic genes will be introduced into cultured cells. Tumor incidence and metastatic potential of expressing clones will be determined in vivo with cells transplanted into the fat pad. Knowledge of the role of individual PKCs in regulation of mammary cell growth is essential for the development of new and alternate strategies for breast cancer treatment and prevention. Specific aims: 1. To compare levels of PKC isozymes and substrates in progressively transformed mammary cells and tumors. 2. To generate mammary cell lines with increased expression of individual PKCs or dominant negative inhibitor PKC constructs. 3. To determine the effects of expressing PKCs and PKC dominant negative inhibitors on tumor incidence and metastasis of transplanted mammary cells in vivo. 4. To generate cell lines with increased expression of two PKC substrates (clones 34 and 72) to assess the role of these proteins in growth and metastasis. 5. To determine the effects of expressing PKCs, PKC dominant negative inhibitors and substrates on 13762 rat mammary cells in culture.