MiRNAs Expression Profiling in Raw264.7 Macrophages after Nfatc1-Knockdown Elucidates Potential Pathways Involved in Osteoclasts Differentiation
Open accessAbstract
Differentiation of macrophages toward osteoclasts is crucial for bone homeostasis but can be detrimental in disease states, including osteoporosis and cancer. Therefore, understanding the osteoclast differentiation process and the underlying regulatory mechanisms may facilitate the identification of new therapeutic targets. Hereby, we tried to reveal new miRNAs potentially involved in the regulation of early steps of osteoclastogenesis, with a particular focus on those possibly correlated with NFATc1 expression, by studying miRNAs profiling. During the first 24 h of osteoclastogenesis, 38 miRNAs were differentially expressed between undifferentiated and RANKL-stimulated RAW264.7 cells, while 10 miRNAs were differentially expressed between RANKL-stimulated cells transfected with negative control or NFATc1-siRNAs. Among others, the expression levels of miR-411, miR-144 and members of miR-29, miR-30, and miR-23 families changed after RANKL stimulation. Moreover, the potential role of miR-124 during osteoclastogenesis was explored by transient cell transfection with anti-miR-124 or miR-124-mimic. Two relatively unknown miRNAs, miR-880-3p and miR-295-3p, were differentially expressed between RANKL-stimulated/wild-type and RANKL-stimulated/NFATc1-silenced cells, suggesting their possible correlation with NFATc1. KEGG enrichment analyses showed that kinase and phosphatase enzymes were among the predicted targets for many of the studied miRNAs. In conclusion, our study provides new data on the potential role and possible targets of new miRNAs during osteoclastogenesis.
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Within the first 24 h of osteoclast formation, the miR-23 family displayed differential downregulation between undifferentiated cells and macrophages stimulated by RANKL.
“Within the first 24 h of osteoclast formation, the miR-23 family displayed differential downregulation between undifferentiated cells and macrophages stimulated by receptor activator of nuclear factor-B ligand (RANKL).”
From Recent advances of miR-23 in human diseases and growth development (2025) · cites this paper as [111]
The Abstract says members of the miR-23 family changed after RANKL stimulation, and Results §3.1 describes a 24-hour comparison of RANKL-stimulated and undifferentiated RAW264.7 cells. Neither passage establishes that the miR-23 family was downregulated in that comparison. The miR-23a-3p downregulation reported in the Discussion instead concerns RANKL-stimulated cells with NFATc1 silenced. (LLM, conf=0.85)
Experimental — accuracy not yet measuredHuman hemoglobin combined with vesicles-derived miR-23a-3p overexpression through gel scaffold can repair and reconstruct bone through the PTEN/AKT signal pathway, and promote osteogenesis and angiogenesis.
“Human hemoglobin combined with vesicles-derived miR-23a-3p overexpression through gel scaffold can repair and reconstruct bone through PTEN/AKT signal pathway, and promote osteogenesis and angiogenesis [[111], [112], [113]].”
From Recent advances of miR-23 in human diseases and growth development (2025) · cites this paper as [111]
The Discussion (page unavailable) reports that miR-23a-3p is highly expressed in exosomes from RANKL-induced RAW264.7 cells and was downregulated after NFATc1 silencing. It does not report a hemoglobin–gel scaffold treatment, bone repair, osteogenesis, angiogenesis, or a PTEN/AKT mechanism. (LLM, conf=0.82)
Experimental — accuracy not yet measuredDuring human cartilage regeneration, expression level of miR-23a-3p in small extracellular vesicles derived from umbilical cord mesenchymal stem cells was significantly ramped up, inhibiting PTEN.
“during human cartilage regeneration, the expression level of miR-23a-3p in small extracellular vesicles derived from umbilical cord mesenchymal stem cells was significantly ramped up, inhibiting the expression level of PTEN.”
From Recent advances of miR-23 in human diseases and growth development (2025) · cites this paper as [111]
The Discussion (page unavailable) discusses miR-23a-3p in exosomes from RANKL-induced RAW264.7 cells and reports its downregulation after NFATc1 silencing. It does not address human cartilage regeneration, vesicles from umbilical-cord mesenchymal stem cells, or PTEN; searches for those terms found no relevant passage in our copy. (LLM, conf=0.77)
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- Authors
- Russo, R., Zito, F., & Lampiasi, N.
- Affiliations
- Institute for Biomedical Research and Innovation (IT)
- Corresponding author
- Lampiasi, Nadia
- Venue
- Biology
- Publisher
- MDPI AG
- Published
- 2021-10-22
- Volume
- 10
- Issue
- 11
- Pages
- 1080
- ISSN
- 2079-7737 (ISSN-L)
- PMC
- PMC8614811
- OpenAlex
- W3208066599
- Cited by
- 6
- References
- 48
- Citations by year
- 2025: 1 · 2024: 4 · 2023: 1
- FWCI
- 0.39
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Biology (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Consiglio Nazionale delle Ricerche (DSB.AD006.146.001)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bone Metabolism and Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
- Health Sciences › Medicine › Rheumatology › Rheumatoid Arthritis Research and Therapies
- Keywords
- microRNA, osteoclastogenesis, NFATc1, RANKL, RAW 264.7 macrophages, miR-124, miR-144, miR-29, miR-30, miR-23, bone homeostasis, osteoporosis
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