急性髓系白血病患者造血干／祖细胞特异性微小RNA的筛选及表达研究  

Screening and expression of CD34＋ cell-specific microRNA in acute myelogenous leukemia

作　　者：王光平[1] 张书芹[1] 朱平[1] 彭敏源[1] 谭三勤[1] 银辉[1] 徐雅静[1] 陈炎[1] 陈方平[1] 

机构地区：[1]中南大学湘雅医院血液科,长沙410008

出　　处：《中华血液学杂志》2012年第7期541-545,共5页Chinese Journal of Hematology

摘　　要：目的筛选和分析急性髓系白血病（AML）患者骨髓造血干／祖细胞（HSPC）特异性微小RNA（miRNA，miR）的表达及其水平。方法分选AML患者骨髓及造血干细胞移植供者（正常对照）外周血CD34＋细胞，提取细胞总RNA，通过与miRNA芯片杂交后筛选差异表达的miRNA，随后采用实时定量PCR技术对差异表达的miR-10a和miR-220c进行定量验证，并进行DNA序列分析。结果miRNA芯片杂交结果显示，AML患者骨髓与正常对照CD34＋细胞之间，表达差异达1倍以上的miRNA有191个。差异有统计学意义的有94个，其中44个表达增高，50个表达降低。实时定量PCR结果显示，AML患者骨髓CD34＋细胞miR-10a和miR-220c表达水平是正常对照的19．6％和19．0％，PCR产物克隆DNA测序以及DNA序列同源检索证实确为miR-10a和miR-220c。结论AML患者与正常人HSPC存在多种差异表达的miRNA，miR-10a和miR-220c表达明显下调。Objective To screen and analyze CD34＋ cell specific microRNAs （miRNAs） from the patients with acute myelogenous leukemia （AML） and their expression. Methods CD34 ＋ cells were sorted from AML patients or the mobilized peripheral blood of the donors of hematopoietic stem cell transplantation （normal control subjects） and followed by the extraction of the cell total RNAs. The differentially expressed microRNAs （miRNAs, miR） were selected after hybridizing with miRNA microarray, real time polymerase chain reaction （real-time PCR） was subsequently applied to confirm the expression of the selected miRs, and PCR products were further cloned and sequenced to check their specificity. Results Of the differentially ex- pressed miRNAs, 191 were found to be at least one-fold change in the CD34 ＋ cells between the AML patients and the normal control subjects. Of the 191 miRNAs, the expression difference of 94 was significant （ P 〈 0.05 ）. Among these 94 miRNAs, the expression of 44 miRNAs was increased and the other 50 miRNAs was decreased in the CD34＋ cells from the bone marrow of AML patients compared with the CD34 ＋ cells from the mobilized peripheral blood of the normal control subjects. Real time PCR verified that the expression level of miR-10a and miR-220c in the CD34＋ cells from the bone marrow of AML patients was 19.6% and 19.0% of that of CD34＋ cells from mobilized peripheral blood of the normal control subjects. DNA sequencing and BLAST DNA database searching results indicated that the PCR products were really miR-10a and miR-220c. Conclusion A variety of differentially expressed-miRNAs are existed between AML and normal control sub- jects CD34 ＋ cells, the expression of miR-10a and miR-220c was significantly down-regulated in the CD34 ＋ cells from the bone marrow of AML patients.

关 键 词：微小RNA 白血病 非淋巴细胞 急性 造血干细胞 

分 类 号：R733.71[医药卫生—肿瘤]