Merkerova Michaela, Belickova Monika, Bruchova Hana
Department of Molecular Genetics, Institute of Hematology and Blood Transfusion, Prague, Czech Republic.
Eur J Haematol. 2008 Oct;81(4):304-10. doi: 10.1111/j.1600-0609.2008.01111.x. Epub 2008 Jun 28.
MicroRNAs (miRNAs) play key roles in a wide variety of normal and pathological cellular processes. A number of studies identified hematopoietic-specific miRNAs that are necessary for correct function of blood cells. Out of our microarray data, we chose 13 miRNAs that showed differential expression in peripheral blood cells (miR-15b, miR-16, miR-24, miR-30c, miR-106b, miR-142-3p, miR-142-5p, miR-150, miR-155, miR-181, miR-223, miR-342, and miR-451) and examined their expression in separated hematopoietic cell lineages.
Using quantitative real-time polymerase chain reaction, we measured relative expression of the miRNAs in fractions of reticulocytes, platelets, granulocytes, monocytes, B- and T-lymphocytes as well as in several hematopoietic cell lines.
We observed that miR-16 and miR-142-3p were highly expressed in all native cell lineages, miR-451 reached the maximal expression in reticulocytes, miR-223 in platelets, granulocytes and monocytes, and miR-150 in B- and T-lymphocytes. Hierarchical clustering analysis grouped the lineage samples according to their origin based on the expression of these miRNAs. To validate discrimination power of the miRNAs, we quantified expression of the 13 miRNAs in several immortalized cell lines. Although the cell lines showed miRNA expression patterns considerably different from those of native cell lineages, clustering analysis distinguished between myeloid, lymphoid and non-hematopoietic cells.
In conclusion, the study reports the expression levels of 13 miRNAs in particular blood cell lineages as well as immortalized cell lines. We demonstrate that the expression profiles of these miRNAs may be used for discrimination of the hematopoietic cell lineages.
微小RNA（miRNA）在多种正常和病理细胞过程中发挥关键作用。多项研究鉴定出对血细胞正常功能至关重要的造血特异性miRNA。从我们的微阵列数据中，我们选择了13种在外周血细胞中显示差异表达的miRNA（miR-15b、miR-16、miR-24、miR-30c、miR-106b、miR-142-3p、miR-142-5p、miR-150、miR-155、miR-181、miR-223、miR-342和miR-451），并检测它们在分离的造血细胞谱系中的表达。
使用定量实时聚合酶链反应，我们测量了miRNA在网织红细胞、血小板、粒细胞、单核细胞、B淋巴细胞和T淋巴细胞亚群以及几种造血细胞系中的相对表达。
我们观察到miR-16和miR-142-3p在所有天然细胞谱系中高表达，miR-451在网织红细胞中表达最高，miR-223在血小板、粒细胞和单核细胞中表达最高，miR-150在B淋巴细胞和T淋巴细胞中表达最高。层次聚类分析根据这些miRNA的表达将谱系样本按其来源进行分组。为了验证miRNA的鉴别能力，我们在几种永生化细胞系中定量了13种miRNA的表达。尽管细胞系显示出与天然细胞谱系有很大不同的miRNA表达模式，但聚类分析区分了髓系、淋巴系和非造血细胞。
总之，该研究报告了13种miRNA在特定血细胞谱系以及永生化细胞系中的表达水平。我们证明这些miRNA的表达谱可用于区分造血细胞谱系。
