A rapid in vitro assay for carcinogenicity of chemical substances in mammalian cells utilizing an attachment‐independence endpoint
A rapid in vitro assay for carcinogenicity of chemical substances in mammalian cells utilizing an attachment‐independence endpoint
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利用不依赖附着的终点对哺乳动物细胞中化学物质的致癌性进行快速体外测定
DOI：
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发表时间：
1979
影响因子：
6.4
通讯作者：
U. Nanna
中科院分区：
文献类型：
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作者：
K. Traul;V. Kachevsky;J. Wolff;R. Hink;U. Nanna
Rauscher白血病病毒（RLV）感染的大鼠胚胎细胞已被证明对多种化学致癌物的肿瘤转化敏感。通常使用的质量测定技术需要6至15周才能观察到形态学改变的细胞灶。虽然软琼脂中的生长被认为是确认肿瘤转化的最佳体外方法之一，但该试验需要在病灶形成后额外4周。我们设计了一种新的快速测定化学品致癌性的方法。该方法利用了RLV感染的大鼠胚胎细胞的化学敏感性范围，但在10天内提供了基于转化细胞在选择附着独立性的条件下存活的能力的读数。用化学品处理靶细胞2FR 450 72小时，然后接种到含有固体底层琼脂/培养基的皮氏培养皿中。在3天和6天后进行活细胞计数，并与溶剂处理的对照细胞的平行数据进行比较。用10种化学致癌物中的任何一种处理2FR 450细胞，包括苯并（a）芘、3-甲基胆蒽、2-乙酰氨基芴、NiSO4和氨基甲酸乙酯等不同化合物，与对照组相比，存活率平均增加225%。非致癌物菲未诱导2FR 450细胞存活率增加。该测定似乎允许在10天内通过简单的测试读数检测各种化学类型的致癌物。
Rauscher leukemia virus (RLV)‐infected rat embryo cells have been shown to be sensitive to neoplastic transformation by a variety of chemical carcinogens. The mass assay technique normally used requires 6 to 15 weeks before morphologically altered foci of cells are observed. While growth in soft agar is considered to be one of the best in vitromeans for confirming neoplastic transformation, this assay requires 4 additional weeks beyond focus formation. We have devised a rapid new assay for determining carcinogenicity of chemicals. The method takes advantage of the range of chemical sensitivity of RLV‐infected rat embryo cells, but provides, in 10 days, for a readout based on the ability of transformed cells to survive under conditions which select for attachment independence. The target cells, 2FR450, were treated for 72 h with chemical and then plated into Petri dishes containing a solid bottom layer of agar/medium. Viable cell counts were made 3 and 6 days later and compared to parallel data from solvent‐treated control cells. 2FR450 cells treated with any of 10 chemical carcinogens, including such diverse compounds as benzo(a)pyrene, 3‐methylcholanthrene, 2‐acetylaminofluorene, NiSO4and urethane, showed an average increase of 225% in survival over controls. Phenanthrene, a non‐carcinogen, induced no increase in survival of 2FR450 cells. This assay appears to permit detection of carcinogens of various chemical types with a simple test readout within 10 days.