目的: 利用基因芯片检测青蒿琥酯作用于K562细胞后的基因表达情况, 从基因水平上探讨青蒿琥酯抑制K562细胞增殖的作用机制。方法: K562细胞经不同浓度青蒿琥酯处理24h, 倒置光显微镜和荧光显微镜观察细胞形态学变化, 流式细胞仪检测细胞周期变化。提取总RNA, 逆转录生成cDNA, cDNA与基因芯片杂交, 扫描仪检测杂交结果。结果: 倒置光显微镜观察到青蒿琥酯处理的K562: 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。荧光显微镜观察到青蒿琥酯处理的K562: 细胞染色质高度浓缩、边缘化, 凝聚成明亮的团块, 即凋亡小体。流式细胞仪检测青蒿琥酯处理K562细胞G₂期细胞的比例明显增加。扫描信号, 分析数据显示10条基因表达有差异, p21、chk1表达上调, cyclinB1、cyclinE1、E2F1、DNA-PK、hTERT、bcl-2、jnk、VEGF表达下调。结论: 青蒿琥酯可以抑制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细胞凋亡有关。

Objective: In order to understand how artesunate inhibited leukaemia cellline K562, on the molecular level, the gene chip was used to detect the gene expression of leukaemia eellline K562 treated by artesunate．Methods: K562 Cells were treated with artesunate for 24h, and then modality changes werestudied by under invert microscope; The morphology changes of the nucleons were observed by Hoechst33342/PI staining; The cell cycle state was examined by flow cytometry(FCM)analysis. Total RNA samples were obtained bv TRIzol and were reversely transcribed to the cDNA．cDNA Samples were hybridized to our gene chips. Hybridization signals were collected and analyzed following scanning by Gene Pix 4100A. Results: The numbers of drift cells were increased and the density of cells was decreased by under invert microscope after K5 62 cells were treated with artesunate for 24h. Morphology changes of cell apoptosis such as karyopyknosis and conglomeration were observed by Hoechst33342/PI staining. FCM Analysis showed that ceils were arrested in G2 phase. There were ten differentially expressed genes identified. Hybridization analysis showed up-regulation of p21 and chkl, and down-regulation of cyclinBl, cyclinEl, E2F1, DNA-PK, hTERT, blc-2，jnk, and VEGF in the artesunate-treated K562 cells. Conclusion: The inhibition of leukaemia eell line K562 by artesunate iS clear：Artesunate exerts its anti-cancer effect by altering the expression of these genes involved in cell cycle and inducing apoptosis.

天津市科委课题资助项目(05YFJMJC08200)