Glutathione-S-transferase A3 knockout mice are sensitive to acute cytotoxic and genotoxic effects of aflatoxin B1.
Glutathione-S-transferase A3 knockout mice are sensitive to acute cytotoxic and genotoxic effects of aflatoxin B1.
复制标题
DOI：
10.1016/j.taap.2009.10.008
复制
发表时间：
2010-02-01
影响因子：
3.8
通讯作者：
Sell S
中科院分区：
文献类型：
--
作者：
Ilic Z;Crawford D;Vakharia D;Egner PA;Sell S
黄曲霉毒素B1（AFB1）是人类肝细胞癌（HCC）的主要危险因素。然而，作为研究AFB1致癌作用的主要动物模型的小鼠，由于小鼠肝脏中谷胱甘肽S-转移酶A3亚基（mGSTA3）的高组成型表达而具有抗性，而人类中缺乏mGSTA3。我们的目的是建立一种小鼠AFB1毒性模型，可用于研究与人类疾病相关的毒性机制，即，一种mGSTA3基因敲除（KO）小鼠，其对AFB1等毒物的反应方式与人类相似。通过同源重组用新霉素盒替换mGSTA3的外显子3 - 6。采用Southern blotting、RT-PCR、Western blotting和AFB1-N7-DNA加合物形成的测定对mGSTA3 KO小鼠进行评价。如预期的，KO小鼠具有mGSTA3基因的外显子3 - 6的缺失，以及在mRNA和蛋白质水平上缺乏mGSTA3表达。注射5 mg/kg AFB1后3小时，mGSTA3 KO小鼠肝脏中的AFB1-N7-DNA加合物比类似治疗的野生型（WT）小鼠多100倍。此外，mGSTA3 KO小鼠在WT小鼠中毒性作用最小的AFB1剂量下死于大面积肝坏死。我们的结论是，mGSTA3基因敲除小鼠对AFB1的急性细胞毒性和遗传毒性作用敏感，证实了GSTA3亚基在保护正常小鼠免受AFB1毒性的关键作用。我们提出mGSTA3基因敲除小鼠作为一个有用的模型，研究导致人类肝癌发展的危险因素的相互作用，以及测试mGSTA3的其他可能的功能。
Aflatoxin B1 (AFB1) is a major risk factor for hepatocellular carcinoma (HCC) in humans. However, mice, the major animal model for the study of AFB1 carcinogenesis, are resistant, due to high constitutive expression, in the mouse liver, of glutathione S-transferase A3 subunit (mGSTA3) that is lacking in humans. Our objective was to establish a mouse model for AFB1 toxicity could be used to study mechanisms of toxicity that are relevant for human disease, i.e., an mGSTA3 knockout (KO) mouse that responds to toxicants such as AFB1 in a manner similar to humans. Exons 3–6 of the mGSTA3 were replaced with a neomycin cassette by homologous recombination. Southern blotting, RT-PCR, Western blotting, and measurement of AFB1-N7-DNA adduct formation were used to evaluate the mGSTA3 KO mice. The KO mice have deletion of exons 3–6 of the mGSTA3 gene, as expected, as well as a lack of mGSTA3 expression at the mRNA and protein levels. Three hours after injection of 5 mg/kg AFB1, mGSTA3 KO mice have more than 100-fold more AFB1-N7-DNA adducts in their livers than do similarly treated wild-type (WT) mice. In addition, the mGSTA3 KO mice die of massive hepatic necrosis, at AFB1 doses that have minimal toxic effects in WT mice. We conclude that mGSTA3 KO mice are sensitive to the acute cytotoxic and genotoxic effects of AFB1, confirming the crucial role of GSTA3 subunit in protection of normal mice against AFB1 toxicity. We propose the mGSTA3 KO mouse as a useful model with which to study the interplay of risk factors leading to HCC development in humans, as well as for testing of additional possible functions of mGSTA3.
登录
查看更多内容
影响因子：
3.9
作者：
Hayes, JD;Chanas, SA;Yamamoto, M
通讯作者：
Yamamoto, M
影响因子：
4.7
作者：
Fields, WR;Morrow, CS;Townsend, AJ
通讯作者：
Townsend, AJ
影响因子：
5.3
作者：
Fernández-Cañón, JM;Baetscher, MW;Grompe, M
通讯作者：
Grompe, M
影响因子：
13.5
作者：
Ghebranious, N;Sell, S
通讯作者：
Sell, S
DOI：
10.1016/j.bbrc.2003.11.068
发表时间：
2003-12-26
影响因子：
3.1
作者：
Jowsey, IR;Smith, SA;Hayes, JD
通讯作者：
Hayes, JD