聚合酶链反应芯片筛选5-氟尿嘧啶对乳鼠心肌细胞心脏毒性差异表达基因的作用  

Polymerase chain reaction array in detecting differentially expressed genes of 5-fluorouracil cardiotoxicity in neonatal rat cardiomyocytes

作　　者：雷和平[1,2] 麦丽萍[1,2] 杨慧[1,2] 钟诗龙[1,2] 杨敏[1,2] 林秋雄[1,2] 单志新[1,2] 余细勇[1,2] 

机构地区：[1]广东省人民医院医学研究中心 [2]广东省医学科学院广东省心血管病研究所,广州510080

出　　处：《岭南心血管病杂志》2014年第3期364-367,共4页South China Journal of Cardiovascular Diseases

基　　金：国家自然科学基金资助项目(项目编号:81202602;81330007;81373486);广东省医学科研基金(项目编号:B2013017)

摘　　要：目的应用聚合酶链反应(polymerase chain reaction,PCR)芯片技术观察5-氟尿嘧啶(5-fluorouracil,5-FU)对乳鼠心肌细胞心脏毒性相关基因表达的调控作用。方法利用差速贴壁法分离培养乳鼠心肌细胞,分为对照组和5-FU刺激组。提取并纯化细胞RNA,紫外分光光度计检测RNA提取物浓度、变性琼脂糖凝胶电泳检测其纯度及完整性。选择包含84个已知心脏毒性相关基因的PCR芯片,采用比较阈值(ΔΔCt)法分析两组基因的表达差异。以表达差异(即上调或下调)大于2倍的基因为有意义的差异基因。结果共有48条基因出现差异表达,其中5条基因表达上调,43条基因表达下调。结论利用PCR芯片技术筛选相关基因,为深入阐明5-FU心脏毒性的作用机制提供了新思路。Objectives To observe the regulatory effect of 5-fluorouracil （5-FU）on related gene expression of cardiact- oxicity in neonatal rat cardiomyocytes by polymerase chain reaction （PCR） array. Methods The cardiomyocytes of neonatal rat were isolated and cultured with differential adhesion method, and were divided into control group and 5-FU stimulation group. After extracting and purifying RNA from the neonatal rat cardiomyocytes, RNA samples were analyzed by ultraviolet spectrophotometer and denaturing agarose gel electrophoresis. Cardiotoxicity PCR array was used to screen the 84 differentially expressed genes, and the data was treated with AACt method. Significant difference was considered when 2- AACt was ≥ 2 or ≤ 0.5. Results Forty-eight genes were detected with differential expressions. Of them, 5 genes were found to be up-regulated and 43 down-regulated. Conclusions PCR array in investigation of differentially expressed genes provides a new approach to further disclose the cardiotoxicity mechanism of 5-FU.

关 键 词：5-氟尿嘧啶 心脏毒性 乳鼠心肌细胞 聚合酶链反应芯片 

分 类 号：R332[医药卫生—人体生理学]