CAJ | 학술논문

　　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。
　　목적관찰방향경수체（AHR）기인침묵후대P19세포향심기분화진정중WNT신호통로적영향。방법근거소서AHR기인서렬설계병구건AHR기인적shRNA진핵표체재체，장AHR간우질립전염지P19세포，통과G418사선출은정적AHR기인침묵표체적양성세포주，응용실시형광정량RT-PCR기술검측P19세포분화과정중WNT신호통로중관건기인GSK3β、β-catenin mRNA적표체수평。결과구건적AHR-shRNA표체질립능현저억제AHR기인적표체；수착P19세포향심기세포분화，실험조β-catenin기인적상대표체량저우대조조，이GSK3β기인적상대표체량칙고우대조조，차이균유통계학의의（P균<0.01）。결론간우AHR기인표체대심장발육과정중WNT신호통로산생조절작용。
Objective To explore the effect of shRNA silenced aromatic hydrocarbon receptor (AHR) on WNT signaling path-way during the differentiation of P19 cells into cardiac myocytes. Methods The eukaryotic expression vector of mouse AHR gene was designed and constructed. The interference plasmid was transfected into P19 cell and the positive stains to AHR gene silencing were screened by G418. The mRNA expression of important genes GSK3βandβ-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 of AHR gene. Along with the differentiation of P19 cell into cardiac myocytes, in the interference group the expression ofβ-catenin gene was lower whereas the expression of GSK3βgene was elevated than those of control group with significant differences (all P<0.01). Conclusions The interference of AHR gene expression can regulate WNT signaling pathway in the development of heart.