IN VITRO SCREENING OF CHEMOPREVENTIVE AGENTS

化学预防剂的体外筛选

基本信息

批准号：
2893918
负责人：
SHEELA SHARMA
金额：
--
依托单位：
ALION SCIENCE AND TECHNOLOGY CORP
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-06-30 至 1999-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2893918
关键词：
benzopyrenes cancer prevention carcinogenesis inhibitor cell transformation chemical carcinogen chemoprevention cytotoxicity drug screening /evaluation histochemistry /cytochemistry laboratory rat retinoate stainings tissue /cell culture

项目摘要

Primary cultures of RTE cells are exposed to carcinogens on Day 1, then morphologically altered epithelial foci are scored at Day 30. Tracheal epithelial cells are isolated from 8 to 12 weeks old male Fisher 344 rats and plated on collagen coated dishes. Initial cytotoxicity tests with the test chemopreventive agent are used to select appropriate concentration ranges for use in the transformation-inhibition assay. From the initial cytotoxicity test, the highest concentration which permits a CFE of 80% or greater compared to control is determined. This concentration and four half- log lower concentrations are included in the chemopreventive test assays. The attached cells are exposed to: media alone, solvent alone, benzo(a)pyrene alone, benzo(a)pyrene plus retinoic acid (or other positive control), or benzo(a)pyrene plus chemopreventive agent. At Day 2 the benzo(a)pyrene is removed and exposure to the chemopreventive agent, retinoic acid (or other positive control), or solvent continued. At 5-7 days three dishes per group are fixed and stained for cytotoxicity determination. At Day 30 all remaining cultures are fixed, stained, and morphologically altered foci scored. The transformation frequency of the various groups are determined. There are 25 chemopreventive agents to be evaluated by this assay.

RTE 细胞的原代培养物在第一天就暴露于致癌物质，然后在第 30 天对形态改变的上皮病灶进行评分。气管上皮细胞分离自 8 至 12 周龄雄性 Fisher 344 只大鼠并铺在胶原涂层培养皿上。最初的使用测试化学预防剂进行细胞毒性测试选择适当的浓度范围用于转化抑制测定。从最初的细胞毒性试验来看，允许 CFE 达到 80% 或更高的最高浓度与对照相比确定。这个浓度和四个半化学预防测试中包含对数较低浓度化验。贴壁细胞暴露于：单独的介质、溶剂单独使用、单独使用苯并(a)芘、使用苯并(a)芘加视黄酸（或其他阳性对照），或苯并（a）芘加化学预防代理人。第 2 天，苯并(a)芘被除去并暴露于化学预防剂、视黄酸（或其他阳性对照），或溶剂继续。 5-7天每组固定三道菜，染色用于细胞毒性测定。第 30 天，所有剩余的培养物被固定、染色，并对形态改变的病灶进行评分。确定各组的转化频率。该测定有 25 种化学预防剂可供评估。