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1、中国实验血液学杂志JournalofExperimentalHematology2010;18(3):559-563559文章编号(ArticleID):1009-2137(2010)03-0559-05论著人白血病细胞株对L门冬酰胺酶敏感性与门冬酰胺合成酶表达水平的相关性李本尚,何映谊,罗长缨,江华,沈树红,蒋黎敏,张蓓,顾龙君上海交通大学医学院上海儿童医学中心血液肿瘤科,上海200127摘要本研究探讨人白血病细胞株门冬酰胺合成酶(asparaginesynthetase,AsnS)的表达水平及与白血病细胞株对LAsparaginase(LAsp)
2、敏感性的关系。用实时定量PCR技术(realtimequantitativePCR,RQPCR)检测Jurkat,HL60,U937,NB4,THP1,Namalwa,Karpas299和K562等8种细胞株基础的和经LAsp处理后的AsnS表达水平,用CCK8实验检测细胞对LAsp处理的敏感性。结果表明,8种细胞株之间AsnS的基础表达水平存在很大程度的变异,经LAsp处理后细胞株的AsnS表达显著上调(p<0.05),AsnS低水平表达细胞对LAsp相对敏感,而AsnS高表达细胞对LAsp相对耐药。在8种细胞株中,U937对LAsp高度敏感,其Asn
3、S表达水平最低,而K562细胞株对LAsp天然耐药,AsnS表达水平最高。结论:AsnS在天门冬氨酸耗竭后的细胞生物学行为中起着重要的作用,AsnS的表达水平反映了细胞对LAsp的敏感性,LAsp在AsnS低表达白血病的治疗中有潜在的应用价值。关键词白血病;门冬酰胺合成酶;左旋门冬酰胺酶;白血病细胞株中图分类号R733.7文献标识码ARelationshipbetweenAsparagineSynthetaseExpressionLevelandCellSensitivitytoLasparaginaseinHumanLeukemicCellLinesLI
4、BenShang,HEYingYi,LUOChangYing,JIANGHua,SHENShuHong,JIANGLiMin,ZhangBei,GULongJunDepartmentofHematology/Oncology,ShanghaiChildrens'MedicalCenter,ShanghaiJiaotongUniversitySchoolofMedicine,Shanghai200127,ChinaCorrespondingAuthor:GULongJun,Professor,SeniorPhysician.Tel:(021)3862616160
5、60,Email:ljguscmc@hotmail.comAbstractThisstudywaspurposedtoexploretherelationshipbetweenasparaginesynthetase(AsnS)mRNAexpressionlevelandthesensitivityofleukemiccelllinestoLasparaginase.TheAsnSmRNAexpressionlevelin8celllines(Jurka,tHL60,U937,NB4,THP1,Namalwa,Karpas299andK562)wasdeterm
6、inedbyrealtimequantitativePCR(RQPCR)basedonfluorescencedyeEvaGreenbeforeandaftertreatmentwithLAsp,andthecellproliferationrateswereanalyzedbyCCK8assay.TheresultsshowedthattherewasasignificantdisparityofAsnSexpressionlevelin8celllines,andthereweresignificantincreasesofAsnSexpressionleveli
7、ncellscoculturedwithLAsp(p<0.05).Ofalltheseeightcelllines,cellssensitivetoLasparaginasehadlowerAsnSexpressionlevelandcellsresistanttoLasparaginasehadhigherAsnSexpression.U937whichwasthemostsensitivetoLasparaginasehadthelowestAsnSexpressionleve,lwhi