黄曲霉毒素B1处理的牛乳腺上皮细胞中STAT5A基因表达的评估
Evaluation of STAT5A Gene Expression in Aflatoxin B1 Treated Bovine Mammary Epithelial Cells.
作者信息
Forouharmehr Ali, Harkinezhad Taher, Qasemi-Panahi Babak
机构信息
Department of Animal Science, Faculty of Agriculture, University of Zanjan, Zanjan, Iran.
出版信息
Adv Pharm Bull. 2013;3(2):461-4. doi: 10.5681/apb.2013.076. Epub 2013 Aug 20.
PURPOSE
Aflatoxin B1 (AFB1) is a potent mycotoxin which has been produced by fungi such as Aspergillus flavus and Aspergillus parasiticus as secondary metabolites due to their growth on food stuffs and induces hepatocellular carcinoma in many animal species, including humans. In the present study, the effect of AFB1 on STAT5A gene expression was investigated in bovine mammary epithelial cells using real time RT-PCR.
METHODS
Bovine mammary epithelial cells were seeded in a 24-well culture plate for three-dimensional (3D) culture in Matrigel matrix. After 21 days of 3D culture and reaching the required number of cells, cells were treated with AFB1 and incubated for 8 h. For real time PCR reaction, total RNA from the cultured and treated cells was extracted and used for complementary DNA synthesis.
RESULTS
The expression of STAT5A gene was significantly down regulated by AFB1 in dose- dependent manner and led to the reduction of proliferation and differentiation of epithelial cells, which has direct effect in milk protein quantity and quality.
CONCLUSION
According to the results, it seems that down regulation of STAT5A gene in AFB1-treated cells maybe due to DNA damage induced by AFB1 in bovine mammary epithelial cells.
目的
黄曲霉毒素B1（AFB1）是一种强效霉菌毒素，由黄曲霉和寄生曲霉等真菌在食品上生长时作为次级代谢产物产生，可在包括人类在内的许多动物物种中诱发肝细胞癌。在本研究中，使用实时逆转录聚合酶链反应（RT-PCR）在牛乳腺上皮细胞中研究了AFB1对信号转导和转录激活因子5A（STAT5A）基因表达的影响。
方法
将牛乳腺上皮细胞接种于24孔培养板中，在基质胶基质中进行三维（3D）培养。三维培养21天且细胞数量达到要求后，用AFB1处理细胞并孵育8小时。对于实时PCR反应，提取培养和处理后细胞的总RNA并用于互补DNA合成。
结果
AFB1以剂量依赖性方式显著下调STAT5A基因的表达，并导致上皮细胞增殖和分化减少，这对乳蛋白的数量和质量有直接影响。
结论
根据结果，似乎AFB1处理的细胞中STAT5A基因的下调可能是由于AFB1在牛乳腺上皮细胞中诱导的DNA损伤。
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