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目的观察8-硝基白杨素(NOChR)抑制人急性髓性白血病细胞系(HL-60)细胞增殖和诱导凋亡的作用,并探讨其作用机制。方法体外培养HL-60细胞;MTT比色法检测增殖活性;流式细胞术定量分析细胞凋亡程度;琼脂糖凝胶电泳观察DNA梯形条带。Western Blot分析HL-60细胞PTEN、P-Akt蛋白表达的影响。结果NOChR可显著抑制HL-60细胞增殖,呈浓度依赖性,IC50为1.34μmol;NOChR具有诱导HL-60细胞凋亡的作用,且呈浓度和时间依赖性。NOChR以浓度和时间依赖方式上调HL-60细胞PTEN蛋白的表达和抑制P-Akt蛋白的表达,其作用可被PPARγ特异性阻断剂GW9662(10μmol)预孵育拮抗。结论NOChR诱导HL-60细胞凋亡作用与其活化PPARγ、上调PTEN蛋白表达、抑制Akt磷酸化相关。
Objective To observe the effect of NOCHR on the proliferation and apoptosis of human acute myeloid leukemia cell line HL-60 and to explore its mechanism. Methods HL-60 cells were cultured in vitro. MTT assay was used to detect the proliferative activity. Flow cytometry was used to analyze the degree of apoptosis. DNA ladder was observed by agarose gel electrophoresis. Western Blot analysis of HL-60 cells PTEN, P-Akt protein expression. Results NOChR significantly inhibited the proliferation of HL-60 cells in a concentration-dependent manner with an IC50 of 1.34 μmol. NOChR had an apoptosis-inducing effect on HL-60 cells in a concentration-and time-dependent manner. NOChR up-regulated the expression of PTEN protein and inhibited the expression of P-Akt protein in HL-60 cells in a time-and concentration-dependent manner, and its effect could be antagonized by the pre-incubation of PPARγ-specific blocking agent GW9662 (10μmol). Conclusion NOChR can induce the apoptosis of HL-60 cells by up-regulating the expression of PTEN and inhibiting the phosphorylation of Akt by activating PPARγ.