Wang Guang-ping, Zhang Shu-qin, Zhu Ping, Peng Min-yuan, Tan San-qin, Yin Hui, Xu Ya-jing, Chen Yan, Chen Fang-ping
Department of Hematology, Xiangya Hospital of Central South University, Changsha 410008, China.
Zhonghua Xue Ye Xue Za Zhi. 2012 Jul;33(7):541-5.
To screen and analyze CD34(+) cell specific microRNAs (miRNAs) from the patients with acute myelogenous leukemia (AML) and their expression.
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.
Of the differentially expressed 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.
A variety of differentially expressed-miRNAs are existed between AML and normal control subjects 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.
筛选并分析急性髓系白血病（AML）患者CD34(+)细胞特异性微小RNA（miRNA）及其表达情况。
从AML患者或造血干细胞移植供者（正常对照者）动员的外周血中分选CD34(+)细胞，随后提取细胞总RNA。与miRNA微阵列杂交后筛选差异表达的微小RNA（miRNA，miR），接着应用实时聚合酶链反应（实时PCR）确认所选miR的表达情况，并对PCR产物进行进一步克隆和测序以检查其特异性。
在差异表达的miRNA中，发现191种在AML患者和正常对照者的CD34(+)细胞之间至少有1倍的变化。在这191种miRNA中，94种的表达差异具有统计学意义（P < 0.05）。在这94种miRNA中，与正常对照者动员外周血的CD34(+)细胞相比，AML患者骨髓CD34(+)细胞中44种miRNA的表达增加，另外50种miRNA的表达降低。实时PCR证实，AML患者骨髓CD34(+)细胞中miR-10a和miR-220c的表达水平分别是正常对照者动员外周血CD34(+)细胞的19.6%和19.0%。DNA测序和BLAST DNA数据库搜索结果表明，PCR产物确实是miR-10a和miR-220c。
AML与正常对照者的CD34(+)细胞之间存在多种差异表达的miRNA，AML患者骨髓CD34(+)细胞中miR-10a和miR-220c的表达明显下调。
