Sox-2基因表达的变化在小鼠EST模型药物胚胎毒性评价中的应用  

Expression of Sox-2 in mouse embryonic stem cell test for screening drugs with embryotoxicity

作　　者：许华[1] 何清[1] 邓淑琴[1] 查庆兵 

机构地区：[1]暨南大学药学院 [2]附属第一医院胎儿医学科,广东广州510632

出　　处：《中国病理生理杂志》2011年第5期985-990,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30973602);广州市科技计划资助项目(No.2010Y1-C851);国家重大新药创制项目(No.2009ZX09103-749)

摘　　要：目的:建立小鼠胚胎干细胞实验(EST)模型,验证该模型检测胚胎毒性的有效性,探讨未分化基因Sox-2表达水平的变化对药物胚胎毒性评价的作用。方法:体外培养小鼠胚胎干细胞(ES),通过形态学观察、核型分析和碱性磷酸酶(AKP)染色等方法鉴定ES;MTT法检测5-氟尿嘧啶(5-FU)、苯妥英钠(DPH)和青霉素G(penicillin G)对ES的毒性作用,RT-PCR半定量分析法检测3种药物对未分化基因Sox-2表达的影响。结果:成功建立EST模型。5-FU、DPH和penicillin G细胞毒性检测结果表明,其胚胎毒性依次为强、弱和无,与临床药物毒性相一致。Sox-2基因表达结果显示,随药物浓度增高及毒性增加,Sox-2基因表达量逐渐高于阴性对照组,但均低于未分化ES细胞。结论:研究结果显示,Sox-2基因的表达水平与药物毒性密切相关,可用于初步评价EST模型中药物的胚胎毒性。AIM： To establish a model of mouse embryonic stem cell test（EST） to examine the toxic effect of 5-fluorouracil（5-FU）,diphenylhydantoin（DPH） and penicillin G on embryonic stem cells and to confirm the validity of the model for detecting the embryotoxicity of drugs.METHODS： Mouse embryonic stem cells were cultured and identified by the methods of morphology,karyotype analysis and alkaline phosphatase staining.The cytotoxic effect of 5-FU,DHP and penicillin G were detected by MTT assay.The effects of 5-FU,DHP and penicillin G on Sox-2 gene expression were determined by semi-quantitative RT-PCR.RESULTS： EST model was successfully established.As the same as the clinical toxicities of the 3 drugs,the embryotoxicity of 5-Fu was strong,that of DPH was weak and no embryotoxicity of penicillin G was observed.With the increase in the concentrations of the drugs,Sox-2 gene expression was gradually higher than that in negative control cells,but still lower than that in undifferentiated ES cells.CONCLUSION： The established EST model can be effectively used for preliminary evaluation of the embryotoxicity of drugs.

关 键 词：模型 小鼠胚胎干细胞 胚胎毒性 有效性 基因 Sox-2 

分 类 号：R991[医药卫生—毒理学]