shRNA沉默芳香烃受体表达对心肌细胞分化过程中WNT信号通路的影响  

Effect of shRNA silenced aromatic hydrocarbon receptor on WNT signaling pathway during the differentiation of cardiac myocytes

作　　者：范志刚[1,2] 朱春[1] 余章斌[1] 胡晓山[1] 李萌萌[1] 朱金改[1] 朱莎莎[1] 韩树萍[1] 

机构地区：[1]南京医科大学附属南京市妇幼保健院儿科,江苏南京210004 [2]马鞍山市妇幼保健院儿科,安徽马鞍山243001

出　　处：《临床儿科杂志》2013年第7期666-669,共4页Journal of Clinical Pediatrics

基　　金：国家自然科学基金资助项目(No.30973213);江苏省医学重点人才基金资助项目(No.RC2011021)

摘　　要：目的观察芳香烃受体(AHR)基因沉默后对P19细胞向心肌分化进程中WNT信号通路的影响。方法根据小鼠AHR基因序列设计并构建AHR基因的shRNA真核表达载体,将AHR干扰质粒转染至P19细胞,通过G418筛选出稳定的AHR基因沉默表达的阳性细胞株,应用实时荧光定量RT-PCR技术检测P19细胞分化过程中WNT信号通路中关键基因GSK3β、β-catenin mRNA的表达水平。结果构建的AHR-shRNA表达质粒能显著抑制AHR基因的表达;随着P19细胞向心肌细胞分化,实验组β-catenin基因的相对表达量低于对照组,而GSK3β基因的相对表达量则高于对照组,差异均有统计学意义(P均<0.01)。结论干扰AHR基因表达对心脏发育过程中WNT信号通路产生调节作用。Objective To explore the effect of shRNA silenced aromatic hydrocarbon receptor （AldR） on WNT signaling path- way during the differentiation of P19 cells into cardiac myocytes. Methods The eukaryotic expression vector ofmouse.4HR gene was designed and constructed. The interference plasmid was transfected into P19 cell and the positive stains to .4HR gene silencing were screened by G418. The mRNA expression of important genes GSK3fl and fl-catenin were evaluated by real-time fluorescent quantita- tive PCR during the differentiation of P19 cells. Results The constructed AHR-shRNA plasmid significantly inhibited the expression ofAHR gene. Along with the differentiation of P19 cell into cardiac myocytes, in the interference group the expression offl-catenin gene was lower whereas the expression of GSK3fl gene was elevated than those of control group with significant differences （all P 〈 0.01）. Conclusions The interference ofAHR gene expression can regulate WNT signaling pathway in the development of heart.

关 键 词：芳香烃受体 心肌细胞分化 WNT信号通路 小鼠 

分 类 号：R725.4[医药卫生—儿科]