Cited by 240MicroRNA in disease regulationGene expression and cancer classificationPlant Molecular Biology Research

The DOI matches its Crossref record; 17 of the 17 references it cites were found in a registry or an index; none of the 19 sources its claims cite is checked for support yet.

Differential expression of microRNAs in hematopoietic cell lineages

Open access
Merkerova, Michaela, Belickova, Monika, Bruchova, Hana·2008·European Journal of Haematology

Abstract

Objectives: 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. Methods: 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. Results: 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. Conclusions: 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.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

This study measured 13 microRNAs by qRT-PCR in separated peripheral blood cell lineages from 10 healthy donors and in immortalized cell lines, identifying lineage-specific expression patterns. It concludes that these miRNA profiles can discriminate hematopoietic cell lineages, although immortalized cell lines showed expression patterns considerably different from native lineages.

Key findings

  • miR-16 and miR-142-3p were highly expressed in most native hematopoietic lineages; miR-451 reached maximal expression in reticulocytes, miR-223 in platelets, granulocytes, and monocytes, and miR-150 in B- and T-lymphocytes.
  • Reticulocytes and platelets showed apparently higher expression across all tested miRNAs (average 10^3 arbitrary units) than other cell types (average 10–10^2 arbitrary units); mean CT values of RNU6B were higher in reticulocytes (31.1 ± 0.4) and platelets (29.8 ± 0.5) than in granulocytes (26.1 ± 0.2), mononuclear cells (27.0 ± 0.2), monocytes (28.0 ± 0.3), B-cells (28.3 ± 0.2), and T-cells (27.6 ± 0.3).
  • Hierarchical clustering grouped lineage samples according to origin; reticulocytes formed the most distinct group, B- and T-lymphocytes clustered together, and most platelets grouped with myeloid cells, though several platelet samples mixed into granulocyte and monocyte clusters.
  • In immortalized cell lines, miR-16 was highly expressed in all samples including non-hematopoietic 293T and HeLa; miR-142 was almost absent in HeLa and 293T; miR-223 was present in myeloid cells; miR-155 was highly expressed only in lymphoblastic UHKT132 and UHKT133; clustering separated non-hematopoietic, myeloid, and lymphoid cell lines.

Methods

  • Peripheral blood was obtained from 10 healthy adult donors (age 25–74 years, median 35; equal male/female distribution). Mononuclear cells and granulocytes were separated by Ficoll-Paque density gradient centrifugation; platelets by centrifugation; reticulocyte RNA from blood lysates by sucrose/KCl centrifugation. Mononuclear cells were further separated into CD3+ T-cells, CD19+ B-cells, and CD14+ monocytes using MACS kits; purity was controlled by FACS and ranged 96.5–99%.
  • Immortalized cell lines included K-562, KU812, MOLM9, ML2, MV4-11, UHKT132, UHKT133, 293T, and HeLa; cells were cultured in RPMI 1640 with 10% fetal bovine serum and antibiotics and harvested after the fourth passage.
  • miRNA expression was measured by qRT-PCR using TaqMan MicroRNA Expression Assays, normalized to endogenous control RNU6B, with relative expression calculated by the delta CT method. Hierarchical clustering used average linkage and Pearson correlation.

Limitations

  • The paper does not include a formal limitations section. The authors state that higher expression levels in reticulocytes and platelets may have been caused by partial disintegration of RNAs not seen by routine techniques, resulting in distinct levels of RNU6B transcript.
  • The authors state that misclassification of some platelet samples in clustering may be due to similarity between platelets and myeloid cells or possible contamination in platelets.
  • Evident from the text: the native cell analysis used 10 healthy donors, and the immortalized cell lines showed miRNA expression patterns considerably different from those of native cell lineages.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1111/j.1600-0609.2008.01111.x

Cites

17 of 17 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Verification

Claims the paper makes

31 Own result · 13 Sources not found · 7 Own interpretation · 2 Own method

Show all 27 claims

  • It may state two separate claims as one. 24
  • Probably not a substantial claim. 8
26 of 26
  • The paper relies on this without support.

    The immortalized cell lines used represent the hematopoietic lineages from which they were derived (myeloid, lymphoid, non-hematopoietic).

    “K-562 has erythrocytic-megakaryocytic origin, KU812 expresses basophilic features, ML2 and MV4-11 originate in monocytes, and UHKT132 and UHKT133 were established from B-lymphoblasts”
  • The paper relies on this without support.

    RNU6B is a stable endogenous control suitable for normalizing miRNA expression across the tested hematopoietic cell lineages and cell lines.

    “The data were normalized to an endogenous control RNU6B (Applied Biosystems).”
    “Since these cells are enucleated with a short lifespan, we may speculate that this observation was caused by partial disintegration of RNAs (not seen by routine techniques) resulting in distinct levels of RNU6B transcript.”
2 of 2

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

1 of 1

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Metadata

Authors
Merkerova, M., Belickova, M., & Bruchova, H.
Venue
European Journal of Haematology
Publisher
Wiley
Published
2008-09-15
Volume
81
Issue
4
Pages
304-310
ISSN
  • 0902-4441 (ISSN-L)
  • 1600-0609
OpenAlex
W2077066570
Cited by
240
References
17
Citations by year
2026: 3 · 2025: 2 · 2024: 4 · 2023: 7 · 2022: 6 · 2021: 10 · 2020: 9 · 2019: 13 · 2018: 10 · 2017: 8 · 2016: 19 · 2015: 14 · 2014: 19 · 2013: 29 · 2012: 25
FWCI
6.92
Type
journal article
Language
en
Open access
Locations
  • European Journal Of Haematology (publishedVersion)
  • PubMed (publishedVersion)
Funders
Ministerstvo Zdravotnictví Ceské Republiky
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene expression and cancer classification
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Molecular Biology Research
Keywords
microRNA, hematopoietic lineage, miR-150, miR-155, miR-142-3p, miR-142-5p, miR-223, miR-451, miR-16, miR-15b, miR-15b-5p, miR-106b, miR-24, miR-30c, miR-342, miR-181, miR-30c-5p, reticulocytes, platelets, granulocytes, monocytes, B cells, T cells, hematopoietic cell lines, gene expression profiling
MeSH terms
Blood Platelets, Cell Differentiation, Gene Expression Regulation, HeLa Cells, Humans, Leukocytes, Reticulocytes, K562 Cells, Oligonucleotide Array Sequence Analysis, Gene Expression Profiling, MicroRNAs, Granulocyte Precursor Cells, Lymphoid Progenitor Cells

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