IN VITRO SCREENING OF POTENTIAL CHEMOPREVENTIVE AGENTS

使用测试化学预防药剂进行的初始生长抑制检测为 用于选择适当的浓度范围以用于 随后进行转化抑制试验。使用五个对数稀释度 从1 mM或在介质或溶剂中的最高可溶浓度 例如无溶剂毒性的二甲基亚砜。原发前列腺 将上皮细胞接种到60 mm的培养皿中，并允许贴壁24小时 几个小时。然后在新鲜的培养基中加入化学预防测试剂， 让培养物孵化72小时，然后所有的培养物都会收到新鲜的 只含化学防腐剂的培养液，再孵化3-5次 几天。在ALL中使用了阳性对照化合物，如维甲酸 化验。然后，将培养物固定并染色。每人三道菜 打分，并确定相对增长。的归纳 人们正在测量人类前列腺上皮细胞的凋亡率。关卡 诱导细胞凋亡的比例正在被测量以回应暴露于 潜在的化学预防药物。五种无毒的浓缩物 测量它们诱导每个药剂的细胞凋亡的能力。这个 前列腺特异性抗原(PSA)的抑制作用正在被测量。一个 前列腺特异性抗原检测被用来定量测量 化学预防性药物对人前列腺癌PSA产生的影响 人前列腺上皮细胞。至少有5种无毒浓度 对每一种化合物进行测试。

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.