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目的:探讨人白血病细胞株K562经青蒿琥酯和青蒿素处理后基因表达的变化及其可能机制。方法:K562细胞经不同浓度青蒿琥酯和青蒿素处理24h后,倒置相差显微镜和荧光显微镜下观察细胞形态学变化,流式细胞仪检测细胞周期变化;提取细胞总RNA,将逆转录生成的cDNA与基因芯片杂交,分析杂交结果。结果:倒置光显微镜:细胞出现不同程度的皱缩,核分裂相减少,细胞密度下降,漂浮细胞增多。荧光显微镜:染色质高度浓缩、边缘化,凝聚成明亮的团块,即凋亡小体。流式细胞仪:G2期细胞的比例明显增加。芯片杂交分析数据,青蒿琥酯处理组有10条基因表达有差异,表达上调的基因有:p21、chk1,表达下调的基因有:cyclinB1、cyclinE1、E2F1、DNA-PK、hTERT、bcl-2、jnk、VEGF;青蒿素处理组有10条基因表达下调:cyclinD1、cdk4、cdk2、cdc2、DNA-PK、DNA-TopoI、mcl-1、erk、jnk、VEGF。结论:青蒿琥酯和青蒿素可以抑制K562细胞增殖,作用机制与改变细胞周期某些调控物质的基因表达、诱导K562细胞凋亡等有关。
Objective: To investigate the changes of gene expression in human leukemia cell line K562 treated with artesunate and artemisinin and its possible mechanism. METHODS: K562 cells were treated with different concentrations of artesunate and artemisinin for 24 h. The morphology of the cells was observed under inverted phase contrast microscopy and fluorescence microscopy. Changes in cell cycle were detected by flow cytometry; total cellular RNA was extracted and reverse transcribed. The cDNA was hybridized with the gene chip and the hybridization result was analyzed. RESULTS: Inverted light microscopy: cells exhibited varying degrees of shrinkage, reduced mitotic phase, decreased cell density, and increased number of floating cells. Fluorescence microscopy: Chromatin is highly concentrated, marginal, and condenses into bright masses, apoptotic bodies. Flow cytometry: The proportion of G2 cells increased significantly. The chip hybridization analysis data showed that there were 10 genes expressed differently in the artesunate-treated group. The up-regulated genes were p21 and chk1. The down-regulated genes were cyclinB1, cyclinE1, E2F1, DNA-PK, hTERT, and bcl-2. , jnk, VEGF; Artemisinin treatment group had 10 genes down-regulated: cyclinD1, cdk4, cdk2, cdc2, DNA-PK, DNA-TopoI, mcl-1, erk, jnk, VEGF. Conclusion: Artesunate and artemisinin can inhibit the proliferation of K562 cells, and the mechanism is related to altering gene expression of certain cell cycle regulators and inducing apoptosis of K562 cells.