Fundamental Toxicological Sciences

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Fundamental Toxicological Sciences
Vol. 3 No. 4 May 31, 2016 p.157-166
Original Article
A high-throughput Bhas 42 cell transformation assay for the determination of the carcinogenicity of three herbal extracts
  • Zhiying Huang (Sun Yat-sen University, China / hzhiying@mail.sysu.edu.cn)
  • Xue Wang (National Centre for Safety Evaluation of Drugs, National Institutes for Food and Drug Control, China / wangxuepaper@sina.com)
Jiang Pu 1) 2) , Ying Wang 1) 3) , Naisong Qi 2) , Wenwan Du 2) , Xiaoyan Tan 2) , Gaofeng Chen 2) , Hairuo Wen 1) , Xin Wang 1) , Yanping Hu 1) , Jie Song 1) , Yuwen Qiu 2) , Jinqiang Liang 2) , Xinlu Fu 2) , Zhiying Huang 2) , Xue Wang 1)
1) National Centre for Safety Evaluation of Drugs, National Institutes for Food and Drug Control, China , 2) Sun Yat-sen University, China , 3) JOINN Laboratories, Inc., China
Keywords: Bhas42, MCA, TPA, High-throughput, Traditional Chinese medicine
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Abstracts

The Bhas 42 cell transformation assay is a sensitive and effective mutation assay, and it has been widely used to detect and evaluate potential carcinogenic risks, including initiation assay and promotion assay. In this transformation assay, the number of transformed foci is the only criterion. However, it is nonquantitative because the shape is irregular. Nevertheless, a high-throughput Bhas 42 cell transformation assay was developed, which includes the usage of hydrogen peroxide in 96-well plates. In this way, transformed foci are quantitatively analysed in proportion to the optical density value (OD value). In this study, the Bhas 42 cell transformation assay was established using the criterion of the OD value. As a result, 3-methylcholanthrene (3-MCA) presented strong tumour initiation activity, whereas the 12-O-tetradecanoylphorbol-13-acetate (TPA) showed strong tumour promotion activity. Lowdaphne Stringbush Flower and Leaf (LSFL, also named as Wikstroemia chamaedaphne (Bunge) Meissn) and Rhubarb (Rheum palmatum L.) showed strong tumour promotion activity, but the tumour promotion activity of Lilac Daphne Flower Bud (LDFB, also named as Daphne genkwa Siebold & Zucc) was equivocal. Surprisingly, the OD value displayed a strong correlation with the number of transformed foci. In conclusion, the high-throughput Bhas 42 cell transformation assay was feasible.

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