哈尔明碱对人白血病HL-60细胞增殖的抑制作用及机制探讨

Inhibition effect and mechanism of harmine on proliferation of human leukemia HL-60 cells

目的探讨哈尔明碱对人白血病HL-60细胞增殖的抑制作用及机制。方法不同浓度哈尔明碱作用于白血病HL-60细胞,MTS法观察哈尔明碱对HL-60细胞增殖的影响,流式细胞仪检测哈尔明碱对HL-60细胞周期的影响,Western blotting、RT-PCR法分别检测哈尔明碱对HL-60细胞STGC3、CDK1、p27、p53、p-AKT蛋白、mRNA表达的变化,报告基因技术考察哈尔明碱对HL-60细胞NF-κB和AP-1转录活性的影响,ELISA技术检测哈尔明碱对HL-60细胞细胞因子TGF-β、IL-1、IL-6和IL-8水平影响。结果哈尔明碱作用后,HL-60细胞增殖被抑制,呈剂量依赖性。1、2μg/mL哈尔明碱作用后,HL-60细胞G0/G1期细胞增多且S期、G2/M期细胞减少,CDK1、p-AKT蛋白表达下降,STGC3、p27和p53蛋白表达升高,CDK1 mRNA表达下降,STGC3、p27、p53 mRNA表达升高,NF-κB和AP-1转录活性下降,细胞因子表达均下降(P均<0.05)。结论哈尔明碱可抑制HL-60细胞的增殖能力,其机制可能与下调CDK1并上调STGC3、p27、p53的表达,抑制细胞TGF-β、IL-1β、IL-6、IL-8等炎症细胞因子的分泌以及NF-κB和AP-1的转录活性有关。

Objective To investigate the inhibition effect and mechanism of harmine on proliferation of human leuke -mia HL-60 cells.Methods Human leukemia HL-60 cells were treated with different concentrations of harmine .The effect of harmine on proliferation of HL-60 cells was observed by MTS assay , and cell cycle was analyzed by flow cytometry . Western blotting and RT-PCR were used to detect the expression changes of STGC 3, CDK1, p27, p53 and p-AKT protein and mRNA, the levels of TGF-β, IL-1, IL-6 and IL-8 were measured simultaneously by ELISA assay , and transcription ac-tivities of NF-κB and AP-1 were detected by reporter gene assay .Results The proliferation of HL-60 cells was inhibited by harmine and in a dose-dependent manner .HL-60 cells were dealed with 1 μg/mL and 2 μg/mL harmine, the cells in G0/G1 phase increased and in S/G2/M phase decreased; the protein expression of CDK 1 and p-AKT was down-regulated significantly, but the protein expression of STGC3, p27 and p53 was up-regulated significantly; the mRNA expression of CDK1 was down-regulated, but the mRNA expression of STGC3, p27 and p53 was up-regulated;and the transcription ac-tivity of NF-κB and AP-1 was decreased , the production of cytokines was decreased .Conclusion Harmine could inhibit the proliferation of HL-60 cells by down-regulating CDK1, down-regulating STGC3, p27 and p53, and inhibiting the levels of TGF-β, IL-1, IL-6 and IL-8 and the transcription activities of NF-κB and AP-1.