青龙衣醇提物对人白血病K562细胞的基因芯片表达的影响  

Effects of Qinglongyi Ethanol Extract on Gene Expression of Human Leukemia K562 Cell

作　　者：张梅[1] 陈永春[2] 

机构地区：[1]黑龙江中医药大学生物化学与分子生物学教研室,黑龙江哈尔滨150040 [2]黑龙江中医药大学解剖教研室,黑龙江哈尔滨150040

出　　处：《中医药导报》2015年第18期20-22,32,共4页Guiding Journal of Traditional Chinese Medicine and Pharmacy

摘　　要：目的:了解青龙衣醇提物对人白血病K562细胞的基因芯片表达的影响,在基因水平探讨青龙衣醇提物抑制K562细胞增殖的作用机制。方法:制备青龙衣醇提物,在37℃恒温、5%CO2培养K562细胞。采用浓度为0.1、0.5、2.5、12.5、62.5 mg/m L的青龙衣醇提物分别处理K562细胞;采用倒置显微镜与荧光显微镜观察处理后K562细胞的形态改变,并采用流式细胞仪检测K562细胞周期。提取细胞的总RNA,生成c DNA,构建基因芯片,并检测基因表达的结果。结果:经过12.5 mg/m L青龙衣醇提物处理后,通过流式细胞仪测定细胞周期:G1期细胞占23.8%,S期占59.9%,G2期占16.3%;而对照组的细胞周期:G1期占22.9%,S期占72.4%,G2期占4.7%;经青龙衣醇提物处理的细胞中,G2期的细胞比例上升,S期细胞比例下降。管家基因为β-actin,构建的基因芯片的特异性、均一性与重复性均良好。表达上调的基因有chk1;下调的基因有cyclin B1、cyclin E1、cyclin D1、cdk4、cdk2、E2F1、PCNA、VEGF、DNA-TopoⅠ、DNA-PK、jnk和mcl-1。结论:青龙衣醇提物通过调控与细胞周期、血管生成、细胞凋亡有关的基因表达,抑制K562细胞的细胞周期进程,从而表现出抗肿瘤作用。Objective： To explore the effects of Qinglongyi ethanol extract on gene expression of human leukemia K562 cell, and to study the mechanism of Qinglongyi ethanol extract inhibited the proliferation of K562 cells at gene level. Method： Qing- longyi ethanol extracts were prepared, K562 cells were cultured in a constant temperature of 37℃, 5% COy K562 cells were treated with Qinglongyi ethanol extracts of concentration of 0.1, 0.5, 2.5, 12.5, 62.5 mg/mL; the morphology of the ceils was ob- served under inverted microscope, fluorescence microscope; cell cycle was tested by flow cytometry. Total cellular RNA was ex- tracted, cDNA was generated, the gene chip was constructed, and the gene expression was detected. Results： After the treatment of 12.5 mg/mL of Qinglongyi ethanol extract, and the cell cycle was determined by flow cytometry： G1 phase cells accounted for 23.8%, 59.9% S phase cells, and 16.3% G2 phase cells; cell cycle of control group： 22.9% G1 phase cells, 72.4% S phase cells, 4.7% G2 phase cells; In Qinglongyi ethanol extract treated cells, the percentage of ceils in G2 phase increased, and the proportion of cells in S phase decreased. Housekeeping genes was ^-actin, gene chips were constructed by 33 genes; Specificity, uniformity and repeatability of the gene chips were good. The up-regulated genes were chkl; the down regulated genes were cyclinB1, cyclinE1, cyclinD1, cdk4, cdk2, E2F1, PCNA, VEGF, DNA-Topo I, BNA-PK, jnk and mcl-1. Conclusions： Qinglongyi ethanol extract can regulate gene expression related to cell cycle, angiogenesis and apoptosis, and it can inhibit cell cycle progression of K562 cells, which showed antitumor effect.

关 键 词：青龙衣醇提物 人白血病K562细胞 基因芯片 细胞周期 

分 类 号：R285.5[医药卫生—中药学]