Matthews E J, Spalding J W, Tennant R W
Center for Food Safety and Applied Nutrition, Food and Drug Administration, Washington, DC 20204.
Environ Health Perspect. 1993 Jul;101 Suppl 2(Suppl 2):319-45. doi: 10.1289/ehp.93101s2319.
This report introduces an improved method of detecting chemical-induced morphological transformation of A-31-1-13 BALB/c-3T3 cells. The new procedure uses an increased target cell population to assess chemical-induced damage by increasing the initial seeding density and by delaying the initiation time of chemical treatment. Furthermore, a newly developed co-culture clonal survival assay was used to select chemical doses for the transformation assay. This assay measured the relative cloning efficiency (RCE) of chemical treatments in high-density cell cultures. In addition, transformation assay sensitivity was enhanced through the use of improved methods to solubilize many chemicals. From a group of 24 chemicals tested in at least two trials, clear evidence of chemical-induced transformation was detected for 12 chemicals (aphidicolin, barium chloride-2H2O, 5-bromo-2'-deoxyuridine, C.I. direct blue 15, trans-cinnamaldehyde, cytosine arabinoside, diphenylnitrosamine, manganese sulfate-H2O, 2-mercaptobenzimidazole, mezerein, riddelliine, and 2,6-xylidine); 2 chemicals had equivocal activity [C.I. direct blue 218 and mono(2-ethylhexyl)phthalate], 9 chemicals were inactive [carisoprodol, chloramphenicol sodium succinate, 4-chloro-2-nitroaniline, C.I. acid red 114, isobutyraldehyde, mono(2-ethylhexyl)adipate, sodium fluoride, and 12-O-tetradecanoylphorbol-13-acetate), and 1 chemical had an indeterminate response (2,6-dinitrotoluene). All positive responses were detected in the absence of an exogenous activation system and exhibited significant activity at two or more consecutive doses. This report also presents a mathematical method that uses t-statistics for rank-ordering the potency of chemical-induced transformation responses. This model detects sensitivity differences in experiments used to evaluate chemical-induced transformation. Furthermore, it provides a method to estimate a chemical's transformation response in terms of the historical behavior of the assay, as well as its future activity. The most active of the 24 chemicals was mezerein, and the least active chemical was diphenylnitrosamine.
本报告介绍了一种改进的检测化学物质诱导A-31-1-13 BALB/c-3T3细胞形态转化的方法。新方法通过提高初始接种密度和延迟化学处理的起始时间，使用增加的靶细胞群体来评估化学物质诱导的损伤。此外，一种新开发的共培养克隆存活试验被用于为转化试验选择化学物质剂量。该试验测量了高密度细胞培养中化学处理的相对克隆效率（RCE）。另外，通过使用改进的方法溶解多种化学物质，提高了转化试验的灵敏度。在至少两项试验中测试的24种化学物质中，有12种化学物质（阿非科林、二水合氯化钡、5-溴-2'-脱氧尿苷、直接蓝15、反式肉桂醛、阿糖胞苷、二苯基亚硝胺、硫酸锰一水合物、2-巯基苯并咪唑、美泽瑞因、瑞香毒素和2,6-二甲基苯胺）检测到化学物质诱导转化的明确证据；2种化学物质具有不确定的活性[直接蓝218和邻苯二甲酸单（2-乙基己基）酯]，9种化学物质无活性[卡立普多、琥珀氯霉素、4-氯-2-硝基苯胺、酸性红114、异丁醛、己二酸单（2-乙基己基）酯、氟化钠和12-O-十四烷酰佛波醇-13-乙酸酯]，1种化学物质有不确定的反应（2,6-二硝基甲苯）。所有阳性反应均在无外源性激活系统的情况下检测到，并且在两个或更多连续剂量下表现出显著活性。本报告还提出了一种使用t统计量对化学物质诱导转化反应的效力进行排序的数学方法。该模型检测用于评估化学物质诱导转化的实验中的灵敏度差异。此外，它提供了一种根据试验的历史行为及其未来活性来估计化学物质转化反应的方法。24种化学物质中活性最强的是美泽瑞因，活性最弱的化学物质是二苯基亚硝胺。
