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不确定的。 生长组分的活性和/或表达水平的变化 与肿瘤相关的调节信号转导通路 已经注意到进展。 理解的功能意义 这些生物化学相关物为开发新的 预防、检测、诊断和治疗疟疾的替代战略 乳腺癌 蛋白激酶C（PKC）是一个广泛表达的蛋白激酶家族， 已知在调节细胞生长的基本机制中重要的酶 和差异化。 PKC也被认为是一种重要的信号转导途径， 由于PKC是肿瘤促进/进展的主要细胞因子， 促进肿瘤的佛波醇酯的受体。 这项建议的目的 是为了提供一个明确的理解PKC同工酶的作用， 在正常乳腺细胞转变为具有 改变生长潜力并随后进入具有侵袭性的细胞， 转移潜能 PKC实际上是一个不同的同工酶家族， 催化磷脂依赖性蛋白磷酸化。 的作用 细胞过程中的单个同工酶尚未确定。 我们 初步数据表明两种PKC同工酶的增加和 两种与癌前病变转化为肿瘤相关的PKC底物 和增加的转移潜能。麦地那和 D. Welch，respectively）. 我们的目标是测试 这些生物化学的关联通过操纵个体的活动 通过表达PKC、PKC显性负性抑制剂和 独特的基质。 将异位基因引入培养的细胞。 表达克隆的肿瘤发生率和转移潜力将被评估。 用移植到脂肪垫中的细胞在体内测定。 知识 单个PKC在调节乳腺细胞生长中作用是 对于开发新的替代乳腺癌治疗策略至关重要 癌症治疗和预防。 具体目标：1. 为了比较 进行性转化乳腺细胞中的PKC同工酶和底物， 肿瘤的 2. 为了产生具有增加的表达的乳腺细胞系， 单个PKC或显性负性抑制剂PKC构建体。 3. 到 确定表达PKC和PKC显性阴性的影响， 抑制移植乳腺细胞的肿瘤发生和转移 in vivo. 4. 产生两种PKC表达增加的细胞系 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. 为了确定表达PKC的影响，PKC 13762大鼠乳腺细胞上的显性负性抑制剂和底物， 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.