摘要
目的 通过反义封闭chk1、chk2基因 ,阻断细胞周期检测点信号传导通路 ,从而增加HL 6 0细胞放疗敏感性。方法 对HL 6 0细胞进行chk1、chk2正义链和反义链的单转染与共转染 ,于转染后 2 4h进行放射线照射 ,照射后 2 4h用Westernblot测量chk1蛋白的变化 ,用流式细胞仪测定细胞周期及细胞凋亡率。结果 反义封闭chk1可增加HL 6 0细胞放疗后凋亡敏感性 ,细胞凋亡率为 2 6 .31%,明显高于正义链的 10 .34%,二者比较差异有显著性 (P <0 .0 5 )。转染chk1反义链的HL 6 0细胞G2 /M期阻滞现象减弱 ,G2 /M期细胞为 38.42 %,转染chk1正义链为 5 4.6 4%,二者比较差异有显著性 (P <0 0 1)。联合反义封闭chk1、chk2具有协同增效作用。结论 反义封闭chk1、chk2基因可增加HL 6 0细胞放疗后凋亡敏感性。
Objective To block signal transduction of cell cycle checkpoints by antisense blocking of chk1/2 gene to increase the radiation sensitivity of HL-60 cell line. Method To transfect the HL-60 cell with chk1/2 antisense and sense chain alone and in combination, expose the cells to irradiation at 24 h after the transfection, the chk1 protein change was assayed by Western blot and the cell cycles and annexinⅤ apoptosis rates by FCM. Results The irradiated apoptosis sensitivity was increased by antisense blocking of chk1 gene in HL-60 cell line, the apoptotic rate was 26.31% being significantly higher than that of the sense blocking (10.34%) (P<0.05), Furthermore, the G 2/M phase blocking phenomenon decreased and a synergic effect was observed in antisense blocking both the chk1 and chk2 genes. Conclusion Antisense blocking of chk1/chk2 could increase the apoptotic sensitivity to irradiation.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2003年第5期253-255,共3页
Chinese Journal of Hematology
基金
国家自然科学基金资助项目 ( 3980 0 14 9)
