POLYAMINE ANALOGS IN ANTI GENE THERAPY FOR BREAST CANCER

描述(改编自《调查者摘要》)：长期目标 为这个项目开发了一种基于三重DNA的反基因策略 乳腺癌的治疗。申请人在上一次资助期间的学习情况 期间表现出显著的结构专一性效应 多胺类似物在稳定三链DNA和促进摄取 乳腺癌细胞中形成三链的寡核苷酸(TFO)。他们的 假设有选择地利用多胺类似物是可行的 提高三链DNA稳定性，增强治疗效果的策略 TFOS的潜力。这一假设将通过以下方式进行检验：1)量化 新型多胺类似物在启动子上稳定三链DNA的能力 用光谱技术测定cMYC、HER2和CyclinD1基因的位点序列， 荧光测量和电泳迁移率改变分析；2) 多胺诱导的TFO缩合在易化中的作用 TFOS的核吸收及TFO积累和稳定性的测定 乳腺癌细胞；3)评价TFOS和TFOS的生物学效应 多胺类似物作为单一药物并通过以下方式联合作用于乳腺癌细胞 靶基因表达的Northern和Western印迹分析，以及多胺 代谢途径；4)TFO/多胺机制的阐明 转录因子对靶点的类似作用。结果： 这些研究将有助于开发一种新的乳房治疗方法 癌症，并促进我们对三链DNA稳定机制的了解 在试管中。

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.