青蒿琥酯和二氢青蒿素抑癌作用机理研究  

Analyzing the mechanisms related withanti-cancer of artesunate using gene chip technology

作　　者：姚丽[1] 何敬和[1] 谢红[1] 呼文亮[1] 陈立军[1] 

机构地区：[1]武警医学院,天津300162

出　　处：《武警医学院学报》2010年第1期13-16,F0004,共5页Acta Academiae Medicinae CPAPF

基　　金：天津市科委资助项目(05YFJMJC08200)

摘　　要：【目的】基因芯片检测青蒿琥酯和二氢青蒿素作用于K562细胞后的基因表达情况,从基因水平上探讨青蒿琥酯和二氢青蒿素抑制K562增殖的作用机制。【方法】K562细胞经不同浓度青蒿琥酯和二氢青蒿素处理,倒置光显微镜和荧光显微镜观察细胞形态变化,流式细胞仪检测细胞周期,MTT检测细胞生长抑制率。提取总RNA,逆转录成cDNA与基因芯片杂交,检测杂交结果。【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。流式细胞仪:G2期细胞的比例增加。MTT检测两种药物对K562细胞生长有抑制作用。扫描杂交信号,分析数据,青蒿琥酯组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;二氢青蒿素组13条基因表达有差异,表达上调的基因有chk1,表达下调的基因有:PCNA、cyclinB1、cyclinD1、cyclinE1、cdk4、cdk2、E2F1、DNA-PK、DNA-TopoⅠ、Mcl-1、jnk、VEGF。【结论】青蒿琥酯和二氢青蒿素抑制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 To study how artesunate and dihydroartemisinin inhibited leukaemia cell line K562,on the molecular level, the gene chip was used to detect the expression panel of genes of leukaemia cell line K562 treated by artesunate. [ Methods ] K562 cells were treated with artesunate and dihydroartemisinin for 24h,and then studied changes under invert microscope;The morphology changes of the nucleons were observed by Hoechst33342/PI staining;The cell cycle state were examined by flow eytometry analysis;determinating inhibitory rate of drugs to K562 cells by MTT.Total RNA samples obtained by TRIzol and were reverse transcribed to the cDNA.cDNA samples were hybridized to our gene chips.Hybridization signal 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 K562 cells were treated artesunate for 24h. Morphology changes of cell apoptosis such as karyopyknosis and conglomeration were observed by Hoechst33342/PI staining.Flow cytometric analysisshowed that cells were arrested in G2 phase, drugs could obviously inhibit proliferation of K562.There were 16 differentially expressed genes identified, up-regulation of p21, chkl and down-regulation of cyclinB1, cyclinE1, E2F1, DNA-PK, hTERT, bcl-2, VEGF, jnk in the artesunate-treated K562 cells,up-regulation of chk1 and down-regulation of PCNA, cyclinBl, cyclinD1, cyclinE1, cdk4, cdk2, E2F1, DNA-PK, DNA-TopoⅠ, MCL-1, ink, VEGF in the dihydroartemisinin-treated K562 cells. [Conclusions ] Inhibition of leukaemia cell line K562 by artesunate and dihydroartemisinin; Artesunate exert its anti-cancer effect by altering the expression of these genes involved in cell cycle; and inducing apoptosis.

关 键 词：基因芯片 青蒿琥酯 二氢青蒿素 细胞周期 细胞凋亡 

分 类 号：R540.4[医药卫生—心血管疾病]