褪黑素联合全反式维甲酸对HL-60细胞作用的研究

Effects of Melatonin and All Trans Retinoic Acid on HL-60 cells

目的观察褪黑素联合全反式维甲酸对人早幼粒细胞白血病细胞株HL-60的增殖、凋亡、分化及细胞周期的影响。方法以1 mmol/L褪黑素与1μmol/L全反式维甲酸联合作用于HL-60细胞,药物作用24 h、48 h、72 h后进行试验。以姬姆萨染色观察细胞形态变化,以CCK-8的方法检测药物的抑制率,以流式细胞术检测细胞凋亡率,细胞膜表面抗原CD11b的表达阳性率,及细胞周期的变化情况。结果褪黑素联合全反式维甲酸作用于HL-60细胞,细胞逐渐出现凋亡,其抑制率24 h、48 h和72h分别为0.31±0.02,0.57±0.06和0.83±0.04,其促进细胞凋亡的比例为15.67±2.38%,26.93±1.60%和50.13±1.76%,其细胞表面CD11b的阳性率为20.98±0.46%,58.96±6.05%和83.80±2.48%;其对细胞周期的影响表现为G1期细胞增多(69.3±0.5%、75.2±0.1%和85.3±0.1%),S期细胞减少(20.1±0.1%、23.2±0.5%和9.2±0.2%),均优于单用褪黑素组或全反式维甲酸组(P<0.05)。结论褪黑素联合全反式维甲酸可以抑制HL-60细胞的增殖、促进其凋亡,诱导其分化,影响其细胞周期,联合用药的作用优于单用褪黑素组或全反式维甲酸组。

Objective To investigate the effects of melatonin（MLT） and all trans retinoic acid（ATRA）on HL-60 cells.Methods The HL-60 cells were treated with 1mmol/L MLT and 1μmol/L ATRA together for different duration（24,48 and 72h） in culture.The morphology of cells was observed after Giemsa dying.The growth of HL-60cells was studied by measurement of CCK-8,while the apoptosis,expression of CD11b and cell cycle were measured by flow cytometry.Results After treated with MLT and ATRA together for 24,48 and 72h,the reduction of the growth of HL-60 cells was 0.31±0.02,0.57±0.06 and 0.83±0.04,the ratio of apoptotic cell death was15.67±2.38%,26.93±1.60% and 50.13±1.76%,the expression of CD11b was 20.98±0.46%,58.96±6.05% and 83.80±2.48%,the ratio of G1 phase cells was 69.3±0.5%、75.2±0.1% and 85.3±0.1%,while S phase cells was 20.1±0.1%、23.2±0.5% and 9.2±0.2%.All results were more obvious than those treated with MLT or ATRA alone（P〈0.05）.Conclusion Treated with MLT and ATRA together,the reduction of the growth of HL-60 cells,the increasing of apoptotic cell death or differentiation,and the inhibition of progression from G1 to S phase of the cell cycle were more obvious than those treated with MLT or ATRA alone.