Effects of mTOR Knockdown by RNA Interference

RNA 干扰造成的 mTOR 敲低效应

基本信息

批准号：
6836869
负责人：
SHARON B CHANG
金额：
$ 4.89万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-01 至 2005-06-17
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Rapamycin, a bacterial macrolide, is emerging as a promising agent for the treatment of cancer. The mammalian Target of Rapamycin (mTOR) controls translation of proteins for growth, proliferation, and progression of the cell cycle. We hypothesize that mTOR may have rapamycin-independent functions as well. Using a model of mTOR knockdown by RNA interference (RNAi) in breast cancer cells, our specific aims are as follows: 1. To determine functional effects of mTOR knockdown and 2. To obtain a comprehensive molecular profile of mTOR knockdown. To address the first aim, we will measure proliferation using cell-viability assays, assess cell size using particle counters, and check apoptosis using caspase and TUNEL assays. To address the second aim, we will use Western analysis to examine known downstream targets of mTOR, and gene-expression arrays to obtain global expression profiles. We anticipate finding significant differences between mTOR knockdown by RNAi and mTOR inhibition by rapamycin, which would lead to further investigation of rapamycin-independent mTOR functions. Such functions may prove to be novel targets in cancer chemotherapy, particularly in the setting of rapamycin-resistance.

描述（由申请人提供）：雷帕霉素是一种细菌大环内酯，正在成为一种有前景的癌症治疗药物。哺乳动物雷帕霉素靶标 (mTOR) 控制蛋白质的翻译，促进细胞周期的生长、增殖和进展。我们假设 mTOR 也可能具有不依赖于雷帕霉素的功能。在乳腺癌细胞中使用 RNA 干扰 (RNAi) 敲低 mTOR 模型，我们的具体目标如下： 1. 确定 mTOR 敲低的功能效应；2. 获得 mTOR 敲低的全面分子谱。为了实现第一个目标，我们将使用细胞活力测定来测量增殖，使用粒子计数器评估细胞大小，并使用 caspase 和 TUNEL 测定检查细胞凋亡。为了实现第二个目标，我们将使用 Western 分析来检查 mTOR 的已知下游靶标，并使用基因表达阵列来获取全局表达谱。我们预计会发现 RNAi 敲低 mTOR 和雷帕霉素抑制 mTOR 之间的显着差异，这将导致进一步研究不依赖雷帕霉素的 mTOR 功能。这些功能可能被证明是癌症化疗的新靶点，特别是在雷帕霉素耐药的情况下。