中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
1期
81-83
,共3页
微小RNA-21%食管癌%反义寡核苷酸%程序性细胞死亡因子4
微小RNA-21%食管癌%反義寡覈苷痠%程序性細胞死亡因子4
미소RNA-21%식관암%반의과핵감산%정서성세포사망인자4
MicroRNA-21%Esophageal cancer%Antisense oligonucleotide%Programmed cell death 4
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.
目的 觀察微小RNA-21 (miRNA-21)反義寡覈苷痠對食管癌EC9706細胞的生長抑製作用.方法 反義寡覈苷痠轉染食管癌EC9706細胞,噻唑藍(MTT)法檢測細胞生長抑製率,實時熒光定量聚閤酶鏈反應(FQ-PCR)檢測細胞miRNA-21水平,流式細胞儀檢測細胞凋亡,FQ-PCR檢測程序性細胞死亡因子4 (PDCD4) mRNA錶達水平,免疫熒光檢測PDCD4蛋白錶達.結果 反義寡覈苷痠抑製細胞活性的最佳濃度為0.5 μmol/L,轉染反義寡覈苷痠後細胞內miRNA-21的錶達水平明顯下調為無義組的0.045 ±0.021,細胞凋亡明顯增加為12.95％,生存率顯著降低為(48.21±9.56)％,差異有統計學意義(P＜0.05).反義寡覈苷痠組中PDCD4 mRNA相對錶達增高為0.452±0.002,且蛋白質錶達水平增高,差異有統計學意義(P＜0.05).結論 miRNA-21反義寡覈苷痠可通過上調PDCD4有效抑製食管癌EC9706細胞增殖併促進凋亡.
목적 관찰미소RNA-21 (miRNA-21)반의과핵감산대식관암EC9706세포적생장억제작용.방법 반의과핵감산전염식관암EC9706세포,새서람(MTT)법검측세포생장억제솔,실시형광정량취합매련반응(FQ-PCR)검측세포miRNA-21수평,류식세포의검측세포조망,FQ-PCR검측정서성세포사망인자4 (PDCD4) mRNA표체수평,면역형광검측PDCD4단백표체.결과 반의과핵감산억제세포활성적최가농도위0.5 μmol/L,전염반의과핵감산후세포내miRNA-21적표체수평명현하조위무의조적0.045 ±0.021,세포조망명현증가위12.95％,생존솔현저강저위(48.21±9.56)％,차이유통계학의의(P＜0.05).반의과핵감산조중PDCD4 mRNA상대표체증고위0.452±0.002,차단백질표체수평증고,차이유통계학의의(P＜0.05).결론 miRNA-21반의과핵감산가통과상조PDCD4유효억제식관암EC9706세포증식병촉진조망.
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.
