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A high-throughput screening method of cell transformation assay in Bhas 42 cells and the research on genistein

中国药理学与毒理学杂志 2013年第3期 576-577页

作者： WANG Ying ZHANG Joe GENG Xing-chao LI Bo WANG Xue National Center for Safety Evaluation of Drugs National Institute for Food and Drug Control Roche Research and Early Development China

OBJECTIVE This study was conducted to establish a high-throughput screening method of cell transformation assay in Bhas 42 cells,aiming to shorten the experimental period and increase the level of objectivity of in vitro cell transformation assay in the detection of carcinogens and to study the transformation potential of *** ① Assay Establishment:Two cell transformation methods were established in our lab for the Bhas 42 cell transformation assay: traditional foci formation method and hydrogen peroxide treatment *** hydrogen peroxide treatment method,transformation was quantified by measuring survival transformed cells OD value at 450 nm using CCK-8 on Day 20 after a treatment of the cell culture for 24 h by hydrogen peroxide on Day *** culture was then fixed,dyed and the number of foci was counted to verify *** the traditional foci formation method,the cell culture was fixed and dyed on Day 21 after plating cells in 96-well plate as described previously.② Study of the transformation effect of genistein in Bhas 42 cells using the H2 O2 treatment ***,a dose range-finding assay was performed to determine genistein concentrations according to the growth of cell treated with genistein in initiation and promotion assays ***,a definitive cell transformation assay was performed using 5 concentrations of genistein in either initiation or promotion *** 96well plate was used for each compound concentration,and each transformation assay also included negative,positive and blank *** ① In the initiation assay with 3-MCA as an initiator using foci formation,the number of wells having transformed foci was 53 wells /plate in positive control and 13 wells /plate in negative *** the promotion assay with TPA as a promoter using foci formation method,the number of wells having transformed foci was 40 wells /plate in positive control and 18 wells /plate in negative *** proportion of wells with tr

关键词： Bhas 42 cell transformation promotive activity non-genotoxic carcinogens genistein

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A high-throughput screening method of cell transformation assay in Bhas 42 cells and the research on genistein

A high-throughput screening method of cell transformation as...

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2013年（第三届）中国药物毒理学年会暨药物非临床安全性评价研究论坛

作者： WANG Ying ZHANG Joe GENG Xing-chao LI Bo WANG Xue National Center for Safety Evaluation of Drugs National Institute for Food and Drug Control Roche Research and Early Development China

OBJECTIVE This study was conducted to establish a high-throughput screening method of cell transformation assay in Bhas 42 cells,aiming to shorten the experimental period and increase the level of objectivity of in vitro cell transformation assay in the detection of carcinogens and to study the transformation potential of ***①Assay Establishment:Two cell transformation methods were established in our lab for the Bhas 42 cell transformation assay: traditional foci formation method and hydrogen peroxide treatment *** hydrogen peroxide treatment method, transformation was quantified by measuring survival transformed cells OD value at 450 nm using CCK-8 on Day 20 after a treatment of the cell culture for 24 h by hydrogen peroxide on Day *** culture was then fixed,dyed and the number of foci was counted to verify *** the traditional foci formation method,the cell culture was fixed and dyed on Day 21 after plating cells in 96-well plate as described previously.②Study of the transformation effect of genistein in Bhas 42 cells using the HO treatment ***,a dose range-finding assay was performed to determine genistein concentrations according to the growth of cell treated with genistein in initiation and promotion assays ***,a definitive cell transformation assay was performed using 5 concentrations of genistein in either initiation or promotion *** 96-well plate was used for each compound concentration,and each transformation assay also included negative,positive and blank ***①In the initiation assay with 3-MCA as an initiator using foci formation,the number of wells having transformed foci was 53 wells/plate in positive control and 13 wells/plate in negative *** the promotion assay with TPA as a promoter using foci formation method,the number of wells having transformed foci was 40 wells/plate in positive control and 18 wells/plate in negative *** proportion of wells with transformed

关键词： Bhas 42 cell transformation promotive activity non-genotoxic carcinogens genistein

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