摘要
目的:初步研究人胚肾细胞HEK293中miRNA-128-1的表达调控机制,为进一步探讨miRNA-128-1在胶质瘤中的表达调控奠定基础。方法:通过软件UCSC预测miRNA-128-1的启动子区域;运用软件UCSC、CBIL预测可能与miRNA-128-1启动子区域结合的候选转录因子;构建相关质粒,包括pB-miRNA-128-1、pShNF1、pShc-Myc、pShTAF1、pShYY1、pShMAF、pShRELA2、pShRNUX1和pShNFkappaB。然后将pB-miRNA-128-1分别与上述质粒共转染人胚肾细胞HEK293,检测报告基因活性,确定候选转录因子的作用性质。结果:共转染pShc-Myc可显著抑制pB-miRNA-128-1启动子报告基因的活性;共转染pShNFkappaB可显著升高pB-miRNA-128-1启动子报告基因的活性;共转染pShNF1、pShTAF1、pSbYY1、pShMAF、pShRELA2或pShRNUX1对pB-miRNA-128-1启动子报告基因的活性未见明显影响。结论:在HEK293细胞miRNA-128-1的表达调控中,c-Myc可能发挥转录激活作用,而NFkappaB可能发挥转录抑制作用。
OBJECTIVE：To discuss mechanism of miRNA-128-1 expression in HEK293 cells and lay the foundation for further study of miRNA-128-1 regulation in glioma.METHODS：Through analyzing UCSC,we found the probable miRNA-128-1 promoter area.We used UCSC and CBIL to obtain the predicted transcription factors binding on the miRNA-128-1 promoter area.Related plasmids were constructed,including pB-miRNA-128-1 promoter, pShNFl,pShc-Myc,pShTAF1,pShYY1,pShMAF,PShRELA2,pShRNUXl and pShNFkappaB.Cotransfecting pB-miRNA-128-1 promoter and pShRNA into HEK293 cells,we analyzed luciferase activity to identify probable transcription factors which could regulate miRNA-128-1 promoter activity.RESULTS：pShc-Myc suppressed the activity of pB-miRNA-128-1 promoter.pShNFkappaB increased the activity of pB-miRNA-128-1 promoter dramatically.pShNF1,pShTAF1,pShYY1,pShMAF,PShRELA2 or pShRNUX1 showed no obvious influence on the expression of pB-miRNA-128-1.CONCLUSION：In HEK 293 cells,c-Myc upregulated whilst NFkappaB downregulated the promoter activity of miRNA-128-1.
出处
《癌变．畸变．突变》
CAS
CSCD
2012年第3期205-208,212,共5页
Carcinogenesis,Teratogenesis & Mutagenesis
