IN VITRO SCREENING OF POTENTIAL CHEMOPREVENTIVE AGENTS

使用测试化学预防剂的初始生长抑制测定是 用于选择合适的浓度范围， 随后转化抑制测定。 使用5个对数稀释度 从1 mM或培养基或溶剂中的最高可溶性浓度 例如二甲亚砜，而没有溶剂毒性。 原发性前列 将上皮细胞接种到60 mm培养皿中，并使其附着24 小时 然后在新鲜培养基中加入化学预防试验剂， 使培养物孵育72小时，然后所有培养物接受新鲜的 仅含有化学预防剂的培养基中，并再孵育3-5 天 阳性对照化合物如类维生素A用于所有实验中。 测定。 然后将培养物固定并染色。 每人三个菜 点进行评分并确定相对生长。 的诱导 在人前列腺上皮细胞中测量细胞凋亡。 水平 细胞凋亡的诱导是通过暴露于 潜在的化学预防剂。 五种无毒浓度正在被 测量每种试剂诱导细胞凋亡的能力。 的 正在测量前列腺特异性抗原（PSA）的抑制。 一 前列腺特异性抗原测定用于定量测量 化学预防剂对前列腺特异性抗原产生的影响 人前列腺上皮细胞 至少5种无毒浓度 对每种化合物进行检测

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