DAPK过表达对HL-60细胞生物学功能及caspase-3表达的影响

Effect of DAPK overexpression on biological behaviors and caspase-3 expression in HL-60 cells

目的探讨死亡相关蛋白激酶(DAPK)过表达对HL-60细胞生物学功能及对caspase-3表达的影响。方法采用RT-PCR技术检测白血病细胞株DAPK基因m RNA的表达。运用真核表达载体p Receiver-M29-DAPK,用脂质体LipofectamineTM2000介导转染HL-60细胞,研究其过表达对白血病细胞凋亡、细胞周期及分化的影响,并对caspase-3表达的影响。结果 DAPK基因在K562、Molt4、U937细胞表达阳性,而HL-60细胞则表达阴性。转染p Receiver-M29-DAPK后,应用流式细胞术和Hoechst33342染色可观察到转染后HL-60细胞出现凋亡,凋亡细胞百分率显著增高。PI单染和瑞氏染色法分析观察转染前后的HL-60细胞,各细胞周期及形态均无改变。转染后caspase-3的表达水平显著增高。结论 DAPK基因过表达的HL-60细胞凋亡增强,但对HL-60细胞周期和细胞分化无影响。Caspase-3可能参与了凋亡调控。

Objective To explore the effect of DAPK overexpression on the biological behaviors and caspase-3 expression in HL-60 cells. Methods The expression of DAPK mRNA was detected by RT-PCR leukemia cell lines K562, Molt4, U937, and HL-60 cells. HL-60 cells were transfected by a eukaryotic expression vector pReceiver-M29-DAPK via LipofectamineTM 2000, and the impact of DAPK overexpression on cell apoptosis, cell cycle, cell differentiation and caspase-3 expression were analyzed. Results DAPK mRNA expression was positive in K562, Molt4 and U937 cells but negative in HL-60 cells. Significantly increased cell apoptosis was observed in pReceiver-M29-DAPK-transfected HL-60 cells by flow cytometry and Hoechst33342 staining. The cell cycle distribution and differentiation showed no significant changes after the transfection. The expression of caspase-3 was significantly increased in the cells after transfection. Conclusion DAPK gene overexpression promotes apoptosis of HL-60 cells without affecting the cell cycle and differentiation. Caspase-3 may be involved in the regulation of cell apoptosis.