小干扰RNA对缺氧PC12细胞中肌细胞增强因子2A蛋白表达的影响  

Small interfering RNA suppresses the increase of myocyte enhancer factor 2A expression in PC12 cells exposed to hypoxia

作　　者：许静[1] 薛梅[1] 陈吉庆[2] 陆超[2] 周国平[2] 周艳[2] 

机构地区：[1]泰州市人民医院药剂科,江苏泰州225300 [2]南京医科大学第一附属医院儿科,江苏南京210029

出　　处：《中国生化药物杂志》2009年第6期400-403,共4页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的研究小干扰RNA(siRNA)对大鼠缺氧PC12细胞中肌细胞增强因子2A(MEF2A)蛋白表达的影响。方法常规培养和缺氧培养PC12细胞。设计合成和筛选针对MEF2A的siRNA序列(MEF2A-siRNA),构建真核细胞表达载体,用脂质体介导转染PC12细胞。实时荧光定量PCR法检测MEF2A的mRNA表达水平。Western blot检测MEF2A蛋白表达量。结果在用含有MEF2A基因的靶向抑制序列MEF2A-siRNA转染后PC12细胞中MEF2A mRNA相对表达量(2-△△CT)为(0.12±0.03),显著低于常规培养的PC12细胞中的表达量(1.01±0.02),抑制率为88.1%(P<0.01)。Western blot检测表明缺氧组中MEF2A蛋白表达量为(98.4±11.7),显著高于正常对照组的(47.5±7.6)(P<0.01)。与缺氧组相比,MEF2A-siRNA+缺氧组中MEF2A蛋白量下调为(59.3±8.4)(P<0.01),表明MEF2A-siRNA转染对缺氧诱导PC12细胞中MEF2A蛋白表达的上调有显著抑制作用。结论缺氧诱导PC12细胞中MEF2A的表达升高,而siRNA转染可以靶向抑制MEF2A基因表达。Purpose To investigate the effect of small interfering RNA on the increase of myocyte enhancer factor 2A（MEF2A） expression in PC12 cells exposed to hypoxia. Methods PC12 cells were cultured under normal conditions or were exposed to hypoxic conditions. Small interfering RNA targeted MEF2A gene （MEF2A-siRNA） was chemically synthesized. Eukaryocytic expression vector was built and transfected into PC12 cells with liposome. The expression of MEF2A mRNA was detected by real-time PCR. Western blot was used to detect the MEF2A protein. Results Compared with normal control（2^-△△CT = 1 .01 ± 0.02）, the mRNA level of MEF2A gene in PC12 cells with the treatment of MEF2A-siRNA was down-regulated significantly by 88% （2^-△△CT = 0.12 ± 0.03, P 〈 0.01 ）. The expression of MEF2A protein in hypoxia-treated PC12 cells was markedly higher than that of normal control（98.4 ± 11.7 and 47.5 ± 7.6, P 〈 0.01） .However, MEF2A-siRNA could significantly suppress the increase of MEF2A protein （P 〈 0.01 ）. Conclusion MEF2A gene silence induced by siRNA might inhibit the increase of MEF2A protein by hypoxia in PC12 cells.

关 键 词：小干扰RNA 肌细胞增强因子2A PC12细胞 

分 类 号：Q78[生物学—分子生物学]