Weight of evidence approach using a TK gene mutation assay with human TK6 cells for follow-up of positive results in Ames tests: a collaborative study by MMS/JEMS

使用人类 TK6 细胞进行 TK 基因突变检测的证据权重法用于跟踪 Ames 试验的阳性结果:MMS/JEMS 的一项合作研究

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作者:Manabu Yasui, Takayuki Fukuda, Akiko Ukai, Jiro Maniwa, Tadashi Imamura, Tsuneo Hashizume, Haruna Yamamoto, Kaori Shibuya, Kazunori Narumi, Yohei Fujiishi, Emiko Okada, Saori Fujishima, Mika Yamamoto, Naoko Otani, Maki Nakamura, Ryoichi Nishimura, Maya Ueda, Masayuki Mishima, Kaori Matsuzaki, Akira

Background

Conflicting

Conclusions

The usefulness of the TK6 assay, by current methods that have not been combined with new technologies such as proteomics, was found to be limited as a follow-up test, although it still may help to reduce some false positive results (20%) in Ames tests. Thus, the combination analysis with toxicoproteomics may be useful for interpreting false positive results raised by 24 h specific reactions in the assay, resulting in the more reduction (> 20%) of false positives in Ames test.

Results

Two of 10 test substances were negative in the overall judgment (20% effective as a follow-up test). Three of these eight positive substances were negative after the short-term treatment and positive after the 24 h treatment, despite identical treatment conditions without S9. A toxicoproteomic analysis of TK6 cells treated with 4-nitroanthranilic acid was thus used to aid the interpretation of the test results. This analysis using differentially expressed proteins after the 24 h treatment indicated that in vitro specific oxidative stress is involved in false positive response in the TK6 assay. Conclusions: The usefulness of the TK6 assay, by current methods that have not been combined with new technologies such as proteomics, was found to be limited as a follow-up test, although it still may help to reduce some false positive results (20%) in Ames tests. Thus, the combination analysis with toxicoproteomics may be useful for interpreting false positive results raised by 24 h specific reactions in the assay, resulting in the more reduction (> 20%) of false positives in Ames test.

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