GENETIC MANIPULATION OF EGR-R EXPRESSION AND RADIOSENSITIVITY OF CARCINOMA CELLS

癌细胞 EGR-R 表达和放射敏感性的基因操纵

• 批准号: 6336435
• 负责人: RUPERT K. SCHMIDT-ULLRICH
• 金额: $ 10.06万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6336435
• 关键词: MCF7 cell; apoptosis; biological signal transduction; breast neoplasms; carcinoma; cell line; cell proliferation; cyclin dependent kinase; epidermal growth factor; gene expression; gene induction /repression; gene therapy; genetic regulation; genetic transduction; growth factor receptors; ionizing radiation; mitogen activated protein kinase; neoplasm /cancer radiation therapy; neoplastic growth; radiation genetics; radiation sensitivity; receptor expression; squamous cell carcinoma; transfection; transforming growth factors

DESCRIPTION: (Applicant's Description) Many human carcinoma cell lines are regulate in their proliferation through autocrine growth regulation involving growth factors, e.g. epidermal growth factor (EGF) or transforming GF-alpha (TGF-alpha), and members of the EGF receptor (EGFR) receptor tyrosine kinases (RTK), Ionizing radiation (IR) in the therapeutic dose range activates EGFR and with repeated exposure induces up-regulation of EGFR, identifying EGFR as early and late IR response gene. A number of downstream components of EGFR-dependent signal transduction pathways are activated by IR including phospholipase C, stimulation of p42/44 MAPkinase, and transcriptional up-regulation of c- Fos. The significance of this activation is indicated by the fact that single or repeated IR exposures can induce a measurable proliferation response in mammary (MCC) and squamous carcinoma cells (SCC), As IR- induced proliferation responses to repeated IR exposures have been implicated in counteracting the toxic effects of IR disruption of this response should result in improve radiotherapeutic outcomes. Disruption of the IR-induced proliferation through genetic modulation of EGFR expression is the overall objective of the proposed experimentation and will accomplished through the following specific aims: (1) Generation of additional MCC and SCC cells expressing inducible mitogenesis-deficient (MUT) or antisense (AS) EGFR and their characterization with respect to growth and expression profile of EGF/TGF-alpha and other erbB RTK; (2) Mechanistic studies on the effects of EGFR-MUT/-AS expression on IR- induced proliferation and stress pathways and their manipulation through Adenovirus (Ad)-mediated transient transfections in order to shift he balance of proliferation and stress responses with one endpoint of apoptosis; (3) Correlation of studies under Specific Aim 2 with overall cellular responses of IR-induced proliferation, clonogenic survival, and apoptotic cell death; (4) As a first step toward the intervention of the IR-induced proliferation responses through transient transfections of EGFR-MUT/-AS, proven under Specific Aim 1-3, using an AD infection system including characterization of those with respect to signal transduction and overall cellular responses.

描述：（申请人的描述）许多人类癌细胞系是 通过自分泌生长调节调节其扩散 涉及生长因素，例如表皮生长因子（EGF）或 转换GF-Alpha（TGF-Alpha）和EGF受体（EGFR）的成员 受体酪氨酸激酶（RTK），电离辐射（IR） 治疗剂量范围激活EGFR，并反复接触诱导 EGFR的上调，将EGFR识别为IR的早期和晚期响应 基因。 EGFR依赖性信号的许多下游组件 通过包括磷脂酶C，包括磷脂酶C的IR激活转导途径 刺激p42/44 mapkinase，以及c-的转录上调 fos。这种激活的重要性表明了以下事实 单一或重复的IR暴露会引起可测量的增殖 乳腺（MCC）和鳞状癌细胞（SCC）的反应， 诱导对重复IR暴露的增生反应已是 涉及抵消IR中断的毒性作用 反应应导致改善放射治疗结果。破坏 IR诱导的通过EGFR表达的遗传调节引起的增殖 是拟议实验的总体目标，将 通过以下特定目的完成：（1）生成 表达诱导有丝分裂缺陷的其他MCC和SCC细胞 （mut）或反义（AS）egfr及其表征 EGF/TGF-Alpha和其他ERBB RTK的生长和表达谱； （2） 关于EGFR-MUT/AS表达对IR-的影响的机理研究 诱导的增殖和压力途径及其通过 腺病毒（AD）介导的瞬态转染以移动 平衡增殖和压力反应与一个端点 凋亡； （3）在特定目标2下的研究与总体 IR诱导的增殖，克隆生存和 凋亡细胞死亡； （4）作为迈向干预的第一步 IR诱导的增殖反应通过瞬时转染 egfr-mut/-as，使用广告感染系统在特定的目标1-3下进行了证明 包括在信号转导方面的表征 和整体细胞反应。