反义微小RNA-21寡核苷酸对食管癌EC9706细胞生长的抑制作用

MicroRNA-21 antisense oligonucleotides to inhibit the growth of EC9706 cells

目的 观察微小RNA-21 （miRNA-21）反义寡核苷酸对食管癌EC9706细胞的生长抑制作用.方法 反义寡核苷酸转染食管癌EC9706细胞,噻唑蓝（MTT）法检测细胞生长抑制率,实时荧光定量聚合酶链反应（FQ-PCR）检测细胞miRNA-21水平,流式细胞仪检测细胞凋亡,FQ-PCR检测程序性细胞死亡因子4 （PDCD4） mRNA表达水平,免疫荧光检测PDCD4蛋白表达.结果 反义寡核苷酸抑制细胞活性的最佳浓度为0.5 μmol/L,转染反义寡核苷酸后细胞内miRNA-21的表达水平明显下调为无义组的0.045 ±0.021,细胞凋亡明显增加为12.95％,生存率显著降低为（48.21±9.56）％,差异有统计学意义（P＜0.05）.反义寡核苷酸组中PDCD4 mRNA相对表达增高为0.452±0.002,且蛋白质表达水平增高,差异有统计学意义（P＜0.05）.结论 miRNA-21反义寡核苷酸可通过上调PDCD4有效抑制食管癌EC9706细胞增殖并促进凋亡.

Objective To observe microRNA （miRNA）-21 antisense oligonucleotide inhibition of the growth of EC9706 cells.Methods Antisense oligonucleotides of EC9706 cells,tetrazolium blue （MTT） assay of cell growth inhibition rate,real-time fluorescent quantitative polymerase chain reaction （FQ-PCR） miRNA-21 level detection cell apoptosis by flow cytometry,FQ-PCR programmed cell death 4 （PDCD4） mRNA expression levels,immunofluorescence PDCD4 protein expression.Results The optimum concentration of antisense oligonucleotide inhibits cell activity was 0.5 μmol/L,after transfection of antisense oligonucleotide expression levels of intracellular miRNA-21 significantly lower for non-defined group of 0.045 ± 0.021,apoptosis a significant increase of 12.95%,the survival rate was significantly lower （48.21 ±9.56）%,the difference was statistically significant （P 〈0.05）.Antisense oligonucleotides group PDCD4 mRNA expression increased relative to 0.452 ± 0.002,and increased protein expression levels,and the difference was statistically significant （P 〈 0.05）.Conclusion miRNA-21 antisense oligonucleotides can increase PDCD4 of EC9706 effectively inhibit cell proliferation and induce apoptosis.