POLYAMINE ANALOGS IN ANTI GENE THERAPY FOR BREAST CANCER

描述（改编自研究者摘要）：长期目标 该项目的目的是开发一种基于三链DNA的抗基因策略， 乳腺癌的治疗申请人在上一次资助期间的学习 期间已经证明了显着的结构特异性效应， 多胺类似物在稳定三链体DNA和促进 乳腺癌细胞中的三链体形成寡核苷酸（TFO）。他们的 假设多胺类似物的选择性利用是可行的， 增加三链体DNA的稳定性和增强治疗性的策略 TFO的潜力。这一假设将通过以下方式进行检验：1）定量 新的多胺类似物在启动子处稳定三链体DNA的能力 通过光谱技术测定cmyc、her 2和cyclinD 1基因的位点序列， 荧光测量和电泳迁移率变动分析; 2） 阐明多胺诱导的TFO缩合在促进 TFO的核摄取以及TFO在细胞中的积累和稳定性的测定 乳腺癌细胞; 3）评估TFO的生物学反应， 多胺类似物作为单一药剂和组合对乳腺癌细胞的作用， 靶基因表达的北方和Western印迹分析，以及多胺 代谢途径;和4）阐明TFO/多胺的机制 类似作用的转录因子的靶位点。结果 这些研究将有助于开发一种新的乳腺癌治疗方法， 癌症，并推进我们对三链DNA稳定化机制的认识 体外

DESCRIPTION (As Adapted From the Investigator's Abstract): The long-term goal for this project is to develop a triplex DNA-based anti-gene strategy for breast cancer treatment. The applicant's studies during the previous funding period have demonstrated a remarkable structural specificity effect of polyamine analogs in stabilizing triplex DNA, and in facilitating the uptake of a triplex forming oligonucleotide (TFO) in breast cancer cells. Their hypothesis is that the selective utilization of polyamine analogs is a viable strategy to increase the stability of triplex DNA, and enhance the therapeutic potential of TFOs. This hypothesis will be tested by: 1) Quantification of the ability of novel polyamine analogs to stabilize triplex DNA at the promoter site sequences of cmyc, her2, and cyclinD1 genes by spectroscopic techniques, fluorescence measurements, and electrophoretic mobility shift assay; 2) elucidation of the role of polyamine -induced TFO condensation in facilitating nuclear uptake of TFOs and determination of TFO accumulation and stability in breast cancer cells; 3) Evaluation of the biologic response of TFOs and polyamine analogs as single agents and in combination on breast cancer cells by Northern and Western blot analysis of target gene expression, and polyamine metabolism pathways; and 4) Elucidation of the mechanism of TFO/polyamine analog action in terms of transcription factors to targeted sites. Results of these studies will help to develop a new therapeutic modality for breast cancer, and advance our knowledge of the mechanism of triplex DNA stabilization in vitro.