青蒿素抗肿瘤作用机制研究

目的:运用基因芯片技术检测青蒿素作用于人红白血病细胞株K562细胞后的基因表达情况,从分子水平探讨青蒿素抑制人红白血病细胞株K562增殖的作用机制。方法:人红白血病细胞株K562细胞经不同浓度青蒿素处理24h,用倒置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,同时Cy3标记,标记好的cDNA与基因芯片杂交,用扫描仪检测杂交结果。结果:倒置显微镜下可见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;荧光显微镜下可观察到染色质高度浓缩、边缘化,凝聚成明亮的团块,即凋亡小体;流式细胞仪检测显示:G2期细胞的比例显著增加;扫描信号分析数据,其中10条基因表达下调:cyclinD1、cdk4、cdk2、cdc2、DNA-PK、DNA-TopoI、mcl-1、erk、jnk、VEGF。结论:青蒿素可抑制人红白血病细胞株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、诱导人红白血病细胞株K562细胞凋亡等有关。

Anti-tumor Mechanism of Artemisinin

OBJECTIVE:To detect the expression of genes of leukemia cell line K562 treated by artemisinin using the gene chip technology and to study the mechanism of artemisinin in the inhibition of leukemia cell line K562 on the molecular level. METHODS:K562 cells were treated with artemisinin for 24h,and then the morphological change of K562 cells were observed under invert microscope and fluorescence microscope. The cell cycle state was examined by flow cytometry analysis (FCM). Total RNA samples were extracted and reverse transcribed to cDNA. Cy3-labelled cDNA samples were hybridized with gene chips.The hybridization results were detected by Gene Pix 4100A. RESULTS: Under invert microscope,different degree of shrinkage of K562 cells was noted,karyoschisis was reduced,cell density was decreased and the numbers of drift cells were increased.Under fluorescence microscope,caryotin was highly concentrated,marginalized and agglomerated to relucent clump,i.e.apoptotic body. Flow cytometric analysis showed that ratio of cells in G2 phase increased markedly. Hybridization analysis showed down-regulation of cyclin D1,cdk4,cdk2,cdc2,DNA-PK,DNA-TopoI,mcl-1,erk,jnk and VEGF in the artemisinin-treated K562 cells.CONCLUSION: The mechanism for artemisinin to inhibit the proliferation of leukemia cell line K562 is related to its action to alter the gene expression of certain regulatory substances involved in cell cycle and induce apoptosis of leukemia cell line K562.