miRNA-223 upregulated by MYOD inhibits myoblast proliferation by repressing IGF2 and facilitates myoblast differentiation by inhibiting ZEB1

Li, Guihuan; Luo, Wen; Abdalla, Bahareldin Ali; Ouyang, Hongjia; Yu, Jin; Hu, Fan; Nie, Qinghua; Zhang, Xiquan

doi:10.1038/cddis.2017.479

Cited by 60 publications

(67 citation statements)

References 51 publications

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“…Western blot was performed as previously described [47]. The following antibodies were used: anti-c-Myc (sc-42, Santa Cruz Biotechnology, CA, USA), anti-p38α (sc-271120, Santa Cruz Biotechnology), anti-p-p38 (sc-166182, Santa Cruz Biotechnology), anti-ERK (sc-376852, Santa Cruz Biotechnology), anti-p-ERK (sc-7383, Santa Cruz Biotechnology), anti-p-JNK (sc-6254, Santa Cruz Biotechnology), anti-Cyclin D1 (BS2436, Bioworld, St Louis Park, MN, USA), anti-Smad2 (ab228807, Abcam, Cambridge, USA), anti-p-Akt1/2/3 (sc-271966, Santa Cruz Biotechnology), anti-Wnt3 (ab50341, Abcam) and anti-Tubulin (BS1482M, Bioworld).…”

Section: Immunoblotting and Immunofluorescencementioning

confidence: 99%

c-Myc inhibits myoblast differentiation and promotes myoblast proliferation and muscle fibre hypertrophy by regulating the expression of its target genes, miRNAs and lincRNAs

et al. 2018

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The transcription factor c-Myc is an important regulator of cellular proliferation, differentiation and embryogenesis. While c-Myc can inhibit myoblast differentiation, the underlying mechanisms remain poorly understood. Here, we found that c-Myc does not only inhibits myoblast differentiation but also promotes myoblast proliferation and muscle fibre hypertrophy. By performing chromatin immunoprecipitation and high-throughput sequencing (ChIP-seq), we identified the genome-wide binding profile of c-Myc in skeletal muscle cells. c-Myc achieves its regulatory effects on myoblast proliferation and differentiation by targeting the cell cycle pathway. Additionally, c-Myc can regulate cell cycle genes by controlling miRNA expression of which dozens of miRNAs can also be regulated directly by c-Myc. Among these c-Myc-associated miRNAs (CAMs), the roles played by c-Myc-induced miRNAs in skeletal muscle cells are similar to those played by c-Myc, whereas c-Myc-repressed miRNAs play roles that are opposite to those played by c-Myc. The cell cycle, ERK-MAPK and Akt-mediated pathways are potential target pathways of the CAMs during myoblast differentiation. Interestingly, we identified four CAMs that can directly bind to the c-Myc 3' UTR and inhibit c-Myc expression, suggesting that a negative feedback loop exists between c-Myc and its target miRNAs during myoblast differentiation. c-Myc also potentially regulates many long intergenic noncoding RNAs (lincRNAs). Linc-2949 and linc-1369 are directly regulated by c-Myc, and both lincRNAs are involved in the regulation of myoblast proliferation and differentiation by competing for the binding of muscle differentiation-related miRNAs. Our findings do not only provide a genome-wide overview of the role the c-Myc plays in skeletal muscle cells but also uncover the mechanism of how c-Myc and its target genes regulate myoblast proliferation and differentiation, and muscle fibre hypertrophy.

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Section: Immunoblotting and Immunofluorescencementioning

confidence: 99%

c-Myc inhibits myoblast differentiation and promotes myoblast proliferation and muscle fibre hypertrophy by regulating the expression of its target genes, miRNAs and lincRNAs

et al. 2018

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“…Studies have shown that miR-223 participates in the regulation of cardiomyocyte glucose metabolism, myoblast proliferation and differentiation, esophageal squamous cell carcinoma and human granulopoiesis [11][12][13]. In the osteogenic differentiation of hBMSCs, miR-223 was found to act as both a positive and a negative regulator, suggesting its potential in the treatment of bone-related diseases [14].…”

Section: Introductionmentioning

confidence: 99%

MicroRNA-223 Suppresses Osteoblast Differentiation by Inhibiting DHRS3

Zhang

Liu

Zheng

et al. 2018

Cell Physiol Biochem

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Background/Aims: In this study, we aimed to use bioinformatics tools to identify the specific miRNAs and mRNAs involved in osteogenic differentiation and to further explore the way in which miRNA regulates osteogenic differentiation. Methods: The microarray GSE80614, which includes data from 3 human mesenchymal stromal cells (hMSCs) and 3 hMSCs after 72 hours (hr) of osteogenic differentiation, was used to screen for differentially expressed mRNAs. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses of these mRNAs were conducted using Gene Set Enrichment Analysis (GSEA). Then, the miRanda website was employed to detect the binding sites of DHRS3. In vitro experiments, including RT-PCR and western blotting, were used to detect miR-233 and DHRS3 expression levels 7 and 14 days (d) after the induction of osteogenic differentiation using human bone marrow-derived mesenchymal stem cells (hBMSCs). The target relationship between miR-223 and DHRS3 was confirmed by a dual luciferase assay. ALP (alkaline phosphatase) staining, ARS (Alizarin Red S) staining and western blotting (Runx2, OPN, OCN) were used to detect the level of osteogenic differentiation after transfection with miR-223 mimics and DHRS3 cDNA. Results: In this study, 127 mRNAs differentially expressed during osteogenic differentiation were identified in GSE80614. GO term and KEGG pathway enrichment analyses found that the retinol metabolism pathway was activated during osteogenic differentiation and that DHRS3, which is involved in the pathway, was upregulated. During osteogenic differentiation in hBMSCs, miR-223 was gradually downregulated, while DHRS3 was upregulated. After 14 days of osteogenic differentiation, ALP and ARS staining assay results showed strong ALP activity and extracellular matrix calcification with the inhibition of miR-223 or the overexpression of DHRS3. Furthermore, the expression levels of Runx2, OPN, and OCN were upregulated with the knockdown of miR-223 or the overexpression of DHRS3, while the simultaneous transfection of a miR-223 agomir and DHRS3 cDNA resulted in no significant difference from the negative control (NC) group. Conclusion: The inhibition of miR-223 promotes the osteogenic differentiation of hBMSCs via the upregulation of DHRS3.

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“…For primary myoblast isolation, at least six embryos at embryo day 11 (E11) were used in each experiment. The sex of each embryos was determined by PCR with the sex-specific primers ( Li et al, 2017 ).…”

Section: Methodsmentioning

confidence: 99%

“…Chicken embryo fibroblast cell line was cultured in high-glucose Dulbecco’s modified Eagle’s medium (Gibco) with 10% fetal bovine serum and 0.2% penicillin/streptomycin. The isolation and culture of chicken primary myoblasts were carried out as previously described ( Li et al, 2017 ).…”

Section: Methodsmentioning

confidence: 99%

TP63 Transcripts Play Opposite Roles in Chicken Skeletal Muscle Differentiation

et al. 2018

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Tumor protein 63 (TP63) comprises multiple isoforms and plays an important role during embryonic development. It has been shown that TP63 knockdown inhibits myogenic differentiation, but which isoform is involved in the underlying myogenic regulation remains uncertain. Here, we found that two transcripts of TP63, namely, TAp63α and ΔNp63α, are expressed in chicken skeletal muscle. These two transcripts have distinct expression patterns and opposite functions in skeletal muscle development. TAp63 has higher expression in skeletal muscle than in other tissues, and its expression is gradually upregulated during chicken primary myoblast differentiation. ΔNp63 can be expressed in multiple tissues and exhibits stable expression during myoblast differentiation. TAp63α overexpression inhibits myoblast proliferation, induces cell cycle arrest, and enhances myoblast differentiation. However, although ΔNp63α has no significant effect on cell proliferation, the overexpression of ΔNp63α inhibits myoblast differentiation. Using isoform-specific overexpression assays following RNA-sequencing, we identified potential downstream genes of TAp63α and ΔNp63α in myoblast. Bioinformatics analyses and experimental verification results showed that the differentially expressed genes (DEGs) between the TAp63α and control groups were enriched in the cell cycle pathway, whereas the DEGs between the ΔNp63α and control groups were enriched in muscle system process, muscle contraction, and myopathy. These findings provide new insights into the function and expression of TP63 during skeletal muscle development, and indicate that one gene may play two opposite roles during a single cellular process.

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miRNA-223 upregulated by MYOD inhibits myoblast proliferation by repressing IGF2 and facilitates myoblast differentiation by inhibiting ZEB1

Cited by 60 publications

References 51 publications

c-Myc inhibits myoblast differentiation and promotes myoblast proliferation and muscle fibre hypertrophy by regulating the expression of its target genes, miRNAs and lincRNAs

c-Myc inhibits myoblast differentiation and promotes myoblast proliferation and muscle fibre hypertrophy by regulating the expression of its target genes, miRNAs and lincRNAs

MicroRNA-223 Suppresses Osteoblast Differentiation by Inhibiting DHRS3

TP63 Transcripts Play Opposite Roles in Chicken Skeletal Muscle Differentiation

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