CAJ | 학술논문

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。
목적：탐토합이명감대인백혈병HL-60세포증식적억제작용급궤제。방법불동농도합이명감작용우백혈병HL-60세포，MTS법관찰합이명감대HL-60세포증식적영향，류식세포의검측합이명감대HL-60세포주기적영향，Western blotting、RT-PCR법분별검측합이명감대HL-60세포STGC3、CDK1、p27、p53、p-AKT단백、mRNA표체적변화，보고기인기술고찰합이명감대HL-60세포NF-κB화AP-1전록활성적영향，ELISA기술검측합이명감대HL-60세포세포인자TGF-β、IL-1、IL-6화IL-8수평영향。결과합이명감작용후，HL-60세포증식피억제，정제량의뢰성。1、2μg/mL합이명감작용후，HL-60세포G0/G1기세포증다차S기、G2/M기세포감소，CDK1、p-AKT단백표체하강，STGC3、p27화p53단백표체승고，CDK1 mRNA표체하강，STGC3、p27、p53 mRNA표체승고，NF-κB화AP-1전록활성하강，세포인자표체균하강（P균＜0．05）。결론합이명감가억제HL-60세포적증식능력，기궤제가능여하조CDK1병상조STGC3、p27、p53적표체，억제세포TGF-β、IL-1β、IL-6、IL-8등염증세포인자적분비이급NF-κB화AP-1적전록활성유관。
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.