Gene-modified embryonic stem cell test to characterize chemical risks.
Gene-modified embryonic stem cell test to characterize chemical risks.
复制标题
基因修饰胚胎干细胞测试可表征化学风险。
DOI：
10.1007/s11356-015-5051-0
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
Morita T.
中科院分区：
文献类型：
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作者：
Kitada K;Kizu A;Teramura T;Takehara T;Hayashi M;Tachibana D;Wanibuchi H;Fukushima S;Koyama M;Yoshida K;Morita T.
使用小鼠胚胎干（ES）细胞进行细胞生长抑制的高通量试验，以评估化学毒性。我们在此使用96孔培养板方法和MTT测定证明，该方法适合于根据化学品的细胞毒性特性对其进行优先排序。为了对化学物质进行分类，我们使用了p53基因修饰的小鼠ES细胞以及野生型ES细胞。p53基因是一种众所周知的肿瘤抑制基因，它控制由DNA损伤剂如烷化剂和辐射引发的程序性细胞死亡（凋亡）和细胞衰老。在本研究中，发现p53缺陷型ES细胞比野生型ES细胞对肿瘤引发剂二乙基亚硝胺（DEN）的抗性更强，提示p53功能障碍抑制了细胞凋亡或衰老。其他肿瘤引发剂，甲基甲磺酸酯（MMS）和N-甲基-N-亚硝基脲（NMU）在野生型和p53缺陷型ES细胞之间的细胞毒性没有明显差异。因此，使用基因修饰的ES细胞的ES测试系统可用于通过检测化学品对细胞凋亡、遗传毒性潜力以及一般细胞毒性的特征影响来对化学品进行分类。
A high-throughput test of cell growth inhibition was performed using mouse embryonic stem (ES) cells to assess chemical toxicities. We herein demonstrated using a 96-well culture plate approach and the MTT assay that this method was suitable for prioritization of chemicals for their cytotoxic properties. In order to categorize chemicals, we usedp53gene-modified mouse ES cells as well aswild-typeES cells. Thep53gene is a well-known tumor suppressor and controls programmed cell death (apoptosis) and cellular senescence that is triggered by DNA-damaging agents such as alkylating agents and radiation. In the present study,p53-deficient ES cells were found to be more resistant to a tumor initiator, diethylnitrosamine (DEN), thanwild-typeES cells, suggesting the inhibition of apoptosis or senescence by a dysfunction inp53.Chromosome aberrations were more frequently detected inp53-deficient ES cells than inwild-typecells, indicating genomic instability due to the deletion ofp53. Other tumor initiators, methyl methanesulfonate (MMS) andN-methyl-N-nitrosourea (NMU), did not reveal apparent differences in cytotoxicity betweenwild-typeandp53-deficient ES cells. Thus, ES test system using gene-modified ES cells may be used to categorize chemicals by detecting their characteristic effects on apoptosis, genotoxic potentials as well as general cytotoxicity.