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1、·中药研究·Δ青蒿素抗肿瘤作用机制研究*呼文亮,姚丽,谢红,陈立军(中国人民武装警察部队医学院,天津市300162)中图分类号R28515;R97911文献标识码A文章编号1001-0408(2008)36-2804-05摘要目的: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cDN
2、A,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
3、达、诱导人红白血病细胞株K562细胞凋亡等有关。关键词基因芯片;青蒿素;细胞周期;细胞凋亡Anti-tumorMechanismofArtemisininHUWen-liang,YAOLi,XIEHong,CHENLi-jun(Dept1ofBiochemistry,MedicalCollegeoftheChinesePeople'sArmedPoliceForces,Tianjin300162,China)ABSTRACTOBJECTIVE:Todetecttheexpressionofgenesofleukemiacelllin
4、eK562treatedbyartemisininusingthegenechiptechnologyandtostudythemechanismofartemisininintheinhibitionofleukemiacelllineK562onthemolecularlevel1METHODS:K562cellsweretreatedwithartemisininfor24h,andthenthemorphologicalchangeofK562cellswereobservedunderinvertmicroscopeand
5、fluorescencemicroscope1Thecellcyclestatewasexaminedbyflowcytometryanalysis(FCM)1TotalRNAsampleswereextractedandreversetranscribedtocDNA1Cy3-labelledcDNAsampleswerehybridizedwithgenechips1ThehybridizationresultsweredetectedbyGenePix4100A1RESULTS:Underinvertmicroscope,di
6、fferentdegreeofshrinkageofK562cellswasnoted,karyoschisiswasreduced,celldensitywasdecreasedandthenumbersofdriftcellswereincreased1Underfluorescencemicroscope,caryotinwashighlyconcentrated,marginalizedandagglomeratedtorelucentclump,i1e1apoptoticbody1Flowcytometricanalysi
7、sshowedthatratioofcellsinG2phaseincreasedmarkedly1Hybridizationanalysisshoweddown-regulationofcyclinD1,cdk4,cdk2,cdc2,DNA-PK,DNA-TopoI,mcl-1,erk,jnkandVEGFintheartemisinin-treatedK562cells1CONCLUSION:Themechanismforartemisinintoinhibittheproliferationofleukemiacellline
8、K562isrelatedtoitsactiontoalterthegeneexpressionofcertainregulatorysubstancesinvolvedincellcycleandinduceapoptosisofl
