CAJ | 학술논문

目的观察微小RNA(miR)-125b在胶质瘤TJ905细胞中与靶基因ERBB2的相互作用.方法通过生物信息学分析发现miR-125b与ERBB2相互配对的区域,合成miR-125b和对照序列,构建含ERBB2的3'UTR野生型和突变型的荧光素酶基因,转染TJ905细胞1×108/L,通过双荧光素酶报告基因检测miR-125b对靶基因ERBB2的3'UTR区的调控作用.转染48 h后提取蛋白,Western blot检测ERBB2的蛋白表达水平.结果生物信息学方法分析显示miR-125b与ERBB2的3'UTR区域存在可能的结合位点.与对照组比较miR-125b inhibitor转染组可以上调ERBB2的mRNA水平是对照组的1.7倍,Western blot法检测ERBB2基因是对照组的1.3倍；而miRNA-mimics组ERBB2的mRNA下降(37.19±3.76)％,Western blot法检测ERBB2基因下降(39.87±7.26)％.结论在胶质瘤TJ905细胞中ERBB2为miR-125b靶基因,miR-125b可以负调节ERBB2的表达.
목적 관찰미소RNA(miR)-125b재효질류TJ905세포중여파기인ERBB2적상호작용.방법 통과생물신식학분석발현miR-125b여ERBB2상호배대적구역,합성miR-125b화대조서렬,구건함ERBB2적3'UTR야생형화돌변형적형광소매기인,전염TJ905세포1×108/L,통과쌍형광소매보고기인검측miR-125b대파기인ERBB2적3'UTR구적조공작용.전염48 h후제취단백,Western blot검측ERBB2적단백표체수평.결과 생물신식학방법분석현시miR-125b여ERBB2적3'UTR구역존재가능적결합위점.여대조조비교miR-125b inhibitor전염조가이상조ERBB2적mRNA수평시대조조적1.7배,Western blot법검측ERBB2기인시대조조적1.3배；이miRNA-mimics조ERBB2적mRNA하강(37.19±3.76)％,Western blot법검측ERBB2기인하강(39.87±7.26)％.결론 재효질류TJ905세포중ERBB2위miR-125b파기인,miR-125b가이부조절ERBB2적표체.
Objective To explore the interaction of microRNA-125b (miR-125b) and its target gene ERBB2 in glioma TJ905 cells.Methods Bioinformatic analysis found that miR-125b and ERBB2 has each other paired regions.We synthesised miR-125b and the control sequence,and constructed containing ERBB2 3' UTR of wild-type and mutant luciferase genes,and then transfected into TJ905 cells.We used the double luciferase reporter gene to analyze the function of miR-125b regulation on the 3'UTR region of the target gene ERBB2.48 h after transfection,protein was extracted,and the ERBB2 protein expression levels were analyzed by Western blotting.Results The bioinformatics analysis showed that miR-125b and ERBB2 3'UTR region has possible binding sites.Compared with the control group,miR-125binhibitor transfection group can up-regulate the expression of ERBB2 mRNA level was 1.7-fold higher than that of control group,the detection of ERBB2 gene Western blotting was about 1.3-fold higher than that of control group,while group niRNA-mimics ERBB2 mRNA fell about (37.19 ± 3.76) ％,the detection of ERBB2 Western blotting gene decreased by (39.87 ± 7.26)％.miR-125b can decline ERBB2 mRNA and protein levels via experimental verification.Conclusion ERBB2 is the target gene of miR-125b in glioma TJ905 cells,and miR-125b can be a negative regulator to ERBB2 expression.