Development and characterization of CHO repair‐proficient cell lines for comparative mutagenicity and metabolism of heterocyclic amines from cooked food
Development and characterization of CHO repair‐proficient cell lines for comparative mutagenicity and metabolism of heterocyclic amines from cooked food
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CHO 修复能力强的细胞系的开发和表征，用于比较熟食中杂环胺的致突变性和代谢
DOI：
10.1002/em.10125
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发表时间：
2003
影响因子：
2.8
通讯作者：
J. Felton
中科院分区：
文献类型：
--
作者：
R. Wu;F. Panteleakos;J. Felton
为了了解修复和代谢在熟食杂环胺致突变性中的作用，我们以前建立了核苷酸切除修复缺陷的CHO 5P3Nat2细胞系，该细胞系共表达小鼠CYP1A2和人N-乙酰转移酶基因。在本研究中，我们用甲烷磺酸乙酯诱变5P3Nat2细胞，并用2-氨基-3-甲基咪唑并[4，5-f]喹啉(IQ)进行细胞毒性筛选，建立了匹配的修复能力细胞系。在差示细胞毒性(DC)实验中，15个克隆中有4个在最高剂量(30μg/ml)的IQ作用下没有细胞毒作用，而修复缺陷的5P3Nat2细胞在0.3μg/ml时表现出100%的细胞毒性。所有四个IQ抗性克隆，其对紫外线的抗性与修复熟练的aa8细胞相似，仍然同时表达CYP1A2和N-乙酰转移酶基因。对5P3Nat2R9克隆的CXPD基因进行序列分析，发现在UV5突变部位发生了A：T→G：C逆转事件。这种碱基的改变导致UV5细胞中116位密码子的酪氨酸在aa8细胞中回复到原来的半胱氨酸，从而恢复野生型CXPD的活性和修复功能。与5P3Nat2细胞相比，修复熟练的5P3Nat2R9细胞株在2-40μg/mlIQ范围内几乎没有表现出IQ诱导的细胞杀伤，并且腺嘌呤磷酸核糖基转移酶(APRT)基因位点的诱导突变水平显著降低。这对匹配的修复熟练/缺乏修复的细胞系不仅可以深入了解杂环胺饮食致癌物如IQ和PhIP的遗传毒性，还可以深入了解核苷酸切除修复对这些化合物最终诱变性的影响。环境。摩尔。诱变剂。41：7-13,2003.©2003 Wiley-Liss公司
In order to understand the role of repair and metabolism in the mutagenicity of heterocyclic amines from cooked foods, we previously developed the nucleotide excision repair‐deficient CHO 5P3NAT2 cell line engineered to coexpress the mouse CYP1A2 and human N‐acetyltransferase genes. In the present study, we have made a matched repair‐competent cell line by mutagenizing 5P3NAT2 cells with ethyl methanesulfonate and selecting for resistance to cytotoxicity by 2‐amino‐3‐methylimidazo[4,5‐f]quinoline (IQ). In the differential cytotoxicity (DC) assay, 4 out of 15 clones showed no cytotoxic effect with IQ at the highest dose (30 μg/ml) tested, in contrast to repair‐deficient 5P3NAT2 cells, which showed ∼ 100% cytotoxicity at 0.3 μg/ml. Subsequently, these IQ‐resistant clones were examined for resistance to killing by UV irradiation. All four IQ‐resistant clones, which show resistance to UV similar to that of repair‐proficient AA8 cells, still express both the CYP1A2 and N‐acetyltransferase genes. Sequence analysis of CXPD cDNA from the 5P3NAT2R9 clone revealed an A:T→G:C reversion event at the site of the UV5 mutation. This base change results in reversion of the codon 116 tyrosine in UV5 cells back to the original cysteine in AA8 cells, thereby restoring wild‐type CXPD activity and repair function. In contrast to 5P3NAT2 cells, the repair‐proficient 5P3NAT2R9 revertant cell line shows little IQ‐induced cell killing, and dramatically lower levels of induced mutation at the adenine phosphoribosyltransferase (Aprt) gene locus over the range of 2–40 μg/ml IQ. This matched pair of repair‐proficient/deficient cell lines can provide insight not only into the genotoxicity of heterocyclic amine dietary carcinogens such as IQ and PhIP, but also into the effects of nucleotide excision repair on the ultimate mutagenicity of these compounds. Environ. Mol. Mutagen. 41:7–13, 2003. © 2003 Wiley‐Liss, Inc.
油炸食品和结肠癌的风险。
影响因子：
5
作者：
Lyon,JL;Mahoney,AW
通讯作者：
Mahoney,AW
兔肝脏中常见蛋白质的单态性和多态性 N-乙酰转移酶活性共存的生化证据。
DOI：
--
发表时间：
1982
期刊：
The Journal of pharmacology and experimental therapeutics
影响因子：
--
作者：
Hein,DW;Hirata,M;Glowinski,IB;Weber,WW
通讯作者：
Weber,WW
修复缺陷仓鼠突变体 UV5 和 UVL-13 CXPD 突变的分子分析。
DOI：
10.1016/0165-7992(94)90012-4
发表时间：
1994
期刊：
Mutation research
影响因子：
--
作者：
Weber,CA;Kirchner,JM;Salazar,EP;Takayama,K
通讯作者：
Takayama,K