vegf反义核酸对hl-60细胞作用和机制研究

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1、•640••论著•中国实脸血液学杂志JournalofExperimentalHematology2004;12(5):640-643文章编号(ArticleID):1009-2137(2004)05-0640-04VEGF反义核酸对HL・60细胞作用和机制研究费嘉，张沮暨南大学医学院血液病研究所，广州510632摘要本研究探讨血管内皮生长因子(VEGF)反叉核酸对HL-60细胞生长的童效和时效关系，并研究其作用机制以揭示VEGF雇因掰的功能)用含20个脱氧核箱核酸的反义核酸A7(用计算机模拟设计和实验筛选出的最优序列)全疏代修

2、饰，以脂质体介导进入细胞，培养72小时后用MTT法计算细胞生长抑制率；采用台盼蓝拒染法每24小时观条细胞存活情况并计数；用Giemsa染色观测细胞形态学改变；用流式细胞仪检测细胞凋亡百分數；用VEGFELISA试剂盒测定蛋白的水平。结果显示，反义药物对HL-60细胞生长有明显抑制作用，呈刑董依赖关系，并且下调VEGF爰白的表达；反义药物对HL-60细胞的凋亡无明显影响。结论：VEGF反义药物抑制HL-60细胞生长的机制是抑制细胞增殖，而不是促进细胞凋亡，内源性VEGF妥白具有促进HL-60细胞增殖的功能，关键词血管内皮生长因子；

3、HL・60细胞；反义寡核昔酸；细胞增殖中图分类号R733.7；R329.28文献标识码AEffectandMechanismofVEGFAntisensePhosphorothioateDeoxynucleotidesonHL-60LeukemicCellsFEIJia,ZHANGHuanInstituteofHematologyfMedicalCollegeofJinanUniversity9Guangzhou510632,ChinaAbstractToexploredose-effectortime-effectofvas

4、cularendothelialgrowthfactor(VEGF)antisensephosphorothioateoligodeoxynucleotides(ASPS-ODN)ongrowthofHL-60cells,andtostudytheeffectmechanismsoastofindnewroleofVEGF,A7twhichwa^themosteffectiveoneofASPS-ODNselectedwithcomputer-aideddesignandexperi・mentalassay,contains20

5、-DNAmodifiedwithphosphorothioateandwastranferedintocellsmediatedwithlipofectin・Afterculturefor72hours,inhibitiverateofcellgrowthwasdetectedwithMTTmethods♦viablecellswerecountedwithtrypanblueexclusioneach24hour,cellconfigurationandapoptosiswereobservedwithGeimsastain

6、ingandflowcytometryrespectively>levelofVEGFproteinwasdetectedwithVEGFELISAkil.Theresultsshowed(hatA7isable(oinhibitcellgrowthofHL-60indose-dependingmannerofASPS»()DN・todown-regulateVEGFproteinexpressionsignificantly•andnottoinduceapoptosisofHL-60cells•Itisconcludedt

7、hat(hereispossibilitythattheinhibitioneffectofVEGFASPS-ODNonHL-60cellgrowthistorestraincellproliferationwithoutinducingapoptosisofUL-60cell.whichwouldinterpretthatendogenousVEGFproteinshaveacapacityofpromotingproliferationofHL-60cell・KeywordsVEGF;HL-60cell;antisense

8、phosphorothioateoligodeoxynudeotides;proliferationJExpHematol2004；12(5):640-643血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)是与血管新