Survivin反义寡核苷酸对k562细胞周期及c-myc的影响

Effect on antisense oligonucleotide targeting survivin in cell cycle and c-myc of k562 cells

目的探讨凋亡抑制基因survivin的反义寡核苷酸对白血病细胞k562细胞周期及cmyc的影响。方法实验分反义组、无义组、脂质体组、空白组,转染后24、48小时流式细胞仪检测细胞周期;免疫组织化学方法检测survivin、cmyc蛋白水平。结果反义寡核苷酸作用后,k562细胞被阻滞在G0/G1期,G0G1期细胞明显增多,S期及G2M期细胞明显减少,细胞周期进程受到明显阻滞,而无义组、脂质体组、空白组细胞周期进程无明显变化(P<0.05);反义寡核苷酸作用24小时后细胞内survivin及cmyc的表达水平下降,但与无义组、脂质体组、空白组比较无显著性差异(P>0.05),而48小时后survivin及cmyc表达水平进一步降低,与各对照组比较有显著性差异(P<0.05)。结论survivin反义寡核苷酸能阻滞K562细胞于G0/G1期、并下调cmyc蛋白的表达。

Objective To study the effect on antisense oligonucleotide targeting survivin, an inhibitor of apoptosis, in cell cycle and c-myc of leukemic cell k562. Methods k562 cells were divided into 4 groups ： ASON group, NSON group, lipofectin group and control group. After thansfection with ASON for 24, 48h, the cell cycle progression was detected by flow cytometry. The protein level of survivin and c-myc were examined by immunohistochemistry. Results After thansfection with ASON, k562 cell were blocked in G0/ G1 phase, the cell number in G0/G1 phase was increased, and the cell number in S and G2/M phase was decreased, the cell cycle progression was inhibited significantly. After thansfection for 48 h , the protein level of survivin and c-myc in ASON group was decreased significantly. Conclusions The cell cycle progression of k562 cells could be inhibited by antisense oligonucleotide targeting survivin and the protein level of c-myc could be downregulated.