应用寡聚核苷酸芯片检测白血病与其同胞供者的白血病基因表达差异  

Identification of Acute Leukemia-Specific Genes from Leukemia Recipient/Sibling Donor Pairs by Distinguishing Study with Oligonucleotide Microarrays

作　　者：孙薏[1] 董陆佳[1] 田方[2] 王升启[3] 贾治林[1] 黄坚[3] 陈泽建[3] 李伍举[3] 陈喜林[2] 朱平[4] 

机构地区：[1]军事医学科学院附属307医院造血干细胞移植中心,北京100039 [2]军事医学科学院附属307医院 [3]军事医学科学院放射医学研究所,北京100850 [4]北京大学医学部第一附属医院血液内科,北京100034

出　　处：《中国实验血液学杂志》2004年第4期450-454,共5页Journal of Experimental Hematology

基　　金：全军医药卫生科研基金 ( 0 1MA0 94) ;国家自然科学基金 ( 3 98890 0 1) ;军事医学科学院创新基金

摘　　要：为了应用寡聚核苷酸基因表达谱芯片研究 9对白血病患者 /同胞供受者对之间的基因表达谱差异以识别主要的疾病相关基因 ,将 16 3条白血病 /肿瘤发病相关基因组群列入芯片设计 ,合成寡核苷酸探针 ,用点样仪点片制成基因芯片。提取病初白血病患者及其供者的骨髓 /外周血标本或其相对应的健康同胞供者经G CSF动员后并经CS 30 0 0细胞分离机采集的浓集的 9份外周血造血干细胞的总RNA ;分别逆转录并进行寡核苷酸探针杂交。结果表明 :与其相应的正常供者配对比较 ,发现 4例ALL患者中存在 6条显著差异表达基因 ,其中上调表达基因 5条(RIZ ,STK 1,T cellleukemia/lymphoma 1A ,Cbp/ p30 0 ,Op18) ,下调表达 1条 (hematopoieticproteoglycancorepro tein) ;5例AML M4和AML M5患者中亦存在 7条显著差异表达基因 ,其中上调表达 6条 (STAT5B ,ligandp6 2fortheLckSH2 ,CST3,LTC4S ,myeloidleukemiafactor 2 ,epb72 ) ,下调表达 1条 (CCR5 )。结论 :选择有高度遗传关联的兄弟姐妹供受者对作为差异研究比照对象 ,利用寡聚核苷酸基因芯片技术进行疾病基因的筛查 ,筛查出了 13条主要异常基因 ;进一步确认了上述基因在白血病分子发病机制中的关键性作用。To explore differentially expressed genes in leukemia gene expression profile and identify main related genes in actue leukemia,gene expression profiles were analyzed in bone marrow/leucopheresis peripheral blood stem cells samples from 9 acute leukemia patients and their sibling donors with the use of oligonucleotide microarrays. 163 reported leukemia-related genes were involved in the study. The oligonucleotide primers were designed,synthesized and spotted on the chemical-material-coated-glass plates in array. The total RNAs were isolated from nine patients′ bone marrow or leucopheresis peripheral blood cells and from nine their sibling donors peripheral blood stem cells treated by G-CSF,then collected by CS-3000 cell selection machine,and were reversely transcribed to cDNAs with the incorporations of fluoresecnt dUTP. the mixed probes were then hybridized to the oligonucleotide microarray. The results showed that in four patient/donor pairs with B-ALL,5 up-regulated (RIZ,STK-1,T-cell leukemia/lymphoma 1A,Cbp/p300,Op18) and 1 down-regulated genes (hematopoietic proteoglycan core protein) were identified;In five patient/donor pairs with AML-M_4 and AML-M_5,6 up-regulated (STAT5B,ligand p62 for the Lck SH2,CST3, LTC4S,myeloid leukemia factor 2 and epb72) and 1 down-regulated genes (CCR5) were identified. In conclusion,on the basis of distinguishing study of specific genetic related recipient/sibling donor pairs,screening leukemia-related genes with oligonucleotide microarrays,a set of 13 up-regulated or down-regulated genes among 163 leukemia-related genes has been identified. The result has further confirmed that above genes play critical role in the molecular mechanism of acute leukemia.

关 键 词：寡聚核苷酸芯片 白血病 供受者对 差异表达基因 造血干细胞移植 

分 类 号：R733.7[医药卫生—肿瘤]