摘要
目的:探讨miR-499体外是否能诱导大鼠骨髓来源的间充质干细胞(MSCs)向心肌细胞分化,并验证其是否通过靶作用于结肠腺瘤样息肉(APC)调控WNT信号通路发挥作用。方法:在MSCs中过表达miR-499后,Q-RT-PCR检测心肌特异性基因GATA4、Nkx2.5、cTnI及miR-499及其靶基因APC的表达情况,双荧光素酶报告系统验证靶基因APC。结果:niR-499、GATA4、Nk2.5、cTnI表达明显上调(P<0.05),但APC表达无明显差异(P=0.3);双荧光素酶报告系统结果显示与转染空载体组比较荧光素酶活性无明显变化(P=0.37);结论:miR-499可促进MSCs向心肌样细胞分化,但不通过靶作用于APC发挥促分化作用。
Objective:To investigate whether the induction of miR-499 differentiates mesenchymal stem cells(MSCs) into cardiomyocytes in vitro,and to verify if these effects were targeted by adenomatous polyps of colon(APC) via mediation of WNT signaling pathway.Methods:Following overexpression of miR-499 in MSCs,the expression of cardiac specific genes GATA4,Nkx2.5,cTnI,miR-499 and the target genes of APC were detected by using Q-RT-PCR.The target gene,APC,was verified by dual luciferase reporter system.Results-.The expression of miR-499,GATA4,Nkx2.5 and cTnI,but not APC(P = 0.30),were significantly up-regulated(all P<0.05).Dual luciferase reporter system showed that luciferase activity of experimental group was not significantly different compared with the empty vector(P=0.31).Conclusion:MiR-499 promotes the differentiation of MSCs into cardiomyocyte-like cells that is not targeted by the APC gene.
出处
《广州医学院学报》
2014年第1期1-4,共4页
Academic Journal of Guangzhou Medical College
基金
国家自然科学基金(31271053)
广东省自然学基金(52013010015314)
广东省中医药局项目(20122098)
