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IN VITRO SCREENING OF POTENTIAL CHEMOPREVENTIVE AGENTS

潜在化学预防剂的体外筛选

基本信息

批准号：
6096052
负责人：
ANN R KENNEDY
金额：
$ 18.72万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-06-30 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/6096052
关键词：
antineoplastics apoptosis bioassay cancer prevention chemoprevention drug screening /evaluation method development prostate specific antigen technology /technique development tissue /cell culture

项目摘要

Initial growth inhibition assays with the test chemopreventive agent are being used to select appropriate concentration ranges for use in the subsequent transformation-inhibition assay. Five log dilutions from 1 mM or the highest soluble concentration in medium or in a solvent such as dimethylsulfoxide without solvent toxicity. Primary prostate epithelial cells shall be seeded into 60 mm dishes and allowed to attach for 24 hours. Then the chemopreventive test agent shall be added in fresh medium and the cultures allowed to incubate 72 hours then all cultures shall receive fresh media containing only the chemopreventive agent and incubated for 3-5 more days. A positive control compound such as a retinoid shall be used in all assays. Then the cultures shall be fixed and stained. Three dishes per point shall be scored and the relative growth determined. The induction of apoptosis in human prostate epithelial cells is being determined. Methodology to quantitatively measure the level of induction of apoptosis in response to exposure to potential chemopreventive agents is being determined. Five nontoxic concentrations are being measured for their ability to induce apoptosis for each agent. A prostate specific antigen assay to quantitatively measure effects of chemopreventive agents on the production of PSA in cultures of human prostate epithelial cells is being perfromed. At least 5 nontoxic concentrations are being tested for each compound.

使用供试化学预防剂进行初始生长抑制试验，以选择用于后续转化抑制试验的适当浓度范围。从1 mM或培养基或溶剂（如二甲基亚砜）中的最高可溶性浓度开始的5个对数稀释度，无溶剂毒性。将原代前列腺上皮细胞接种到60 mm培养皿中，并使其附着24小时。然后，将化学预防试验试剂加入新鲜培养基中，并使培养物孵育72小时，然后所有培养物应接受仅含化学预防试剂的新鲜培养基，并再孵育3-5天。在所有试验中应使用阳性对照化合物，如类维生素A。然后将培养物固定并染色。每个点应评分三个培养皿，并确定相对生长。人前列腺上皮细胞凋亡的诱导正在确定中。正在确定定量测量响应于暴露于潜在化学预防剂的细胞凋亡诱导水平的方法。正在测量五种无毒浓度的每种药物诱导细胞凋亡的能力。正在进行前列腺特异性抗原测定，以定量测量化学预防剂对人前列腺上皮细胞培养物中PSA产生的影响。每种化合物至少测试5种无毒浓度。