# miRNA-182-5p promotes myogenic differentiation of C2C12 cells via the suppression of ZBTB7A

**Authors:** Mengyuan Zhang, Yongheng Wang, Shan Shan, Siyu Liu

PMC · DOI: 10.3389/fvets.2025.1637277 · Frontiers in Veterinary Science · 2025-07-30

## TL;DR

This study shows that miR-182-5p helps muscle cells differentiate by reducing the activity of ZBTB7A, a protein that may hinder this process.

## Contribution

The study identifies ZBTB7A as a direct target of miR-182-5p in promoting myogenic differentiation.

## Key findings

- Overexpression of miR-182-5p enhances myogenic differentiation of C2C12 cells.
- ZBTB7A is a direct target of miR-182-5p during myogenic differentiation.
- Inhibition of miR-182-5p reduces the differentiation ability of C2C12 cells.

## Abstract

Skeletal muscle possesses a significant regenerative capacity, which is largely mediated by myogenic satellite stem cells. MicroRNAs are known regulators of muscle development. miR-182-5p plays important roles in cell proliferation and migration in various cell types and pathologies. However, its specific role in myogenesis remains unclear. In this study, we elucidated the function of miR-182-5p in the differentiation of C2C12 myoblasts.

We evaluated the effects of overexpression and inhibition of miR-182-5p in C2C12 cells on its myogenic differentiation ability using Giemsa staining. We also determined the mRNA and protein levels of myogenic differentiation marker genes in these cells at different time points after the induction of differentiation in these cells. The target of miR-182-5p was predicted using bioinformatics tools and validated using luciferase reporter assay.

Overexpression of miR-182-5p via mimic transfection promoted differentiation, while its inhibition by a specific compound attenuated this process. Furthermore, using bioinformatic prediction and validation via a dual-luciferase reporter assay, we identified zinc finger and BTB domain containing 7A (Zbtb7a) as a direct target gene of miR-182-5p during C2C12 myogenic differentiation.

Our findings indicate that miR-182-5p positively regulates C2C12 differentiation, partly via the suppression of Zbtb7a and suggest that appropriate miR-182-5p expression is essential for normal myogenesis.

## Linked entities

- **Genes:** ZBTB7A (zinc finger and BTB domain containing 7A) [NCBI Gene 51341]
- **Proteins:** ZBTB7A (zinc finger and BTB domain containing 7A)

## Full-text entities

- **Genes:** Jmjd1c (jumonji domain containing 1C) [NCBI Gene 108829] {aka 5430433L24Rik, D630035I23Rik, Jmjdic, TRIP8}, LOC536229 (paired box protein Pax-7) [NCBI Gene 536229] {aka PAX7}, Tns3 (tensin 3) [NCBI Gene 319939] {aka F830010I22Rik, TEM6, Tens1}, Myh4 (myosin, heavy polypeptide 4, skeletal muscle) [NCBI Gene 17884] {aka MHC2B, MM, MYH-2B, Minimsc, Minmus, MyHC-IIb}, CFL2 (cofilin 2) [NCBI Gene 539332], MYOD1 (myogenic differentiation 1) [NCBI Gene 281938] {aka MyoD}, Rb1 (RB transcriptional corepressor 1) [NCBI Gene 19645] {aka Rb, Rb-1, p110-RB1, pRb, pp105}, MYOG (myogenin) [NCBI Gene 281343], Mir26a-1 (microRNA 26a-1) [NCBI Gene 387218] {aka Mirn26a, Mirn26a-1, miR-26a, mir-26a-1}, Dab2ip (disabled 2 interacting protein) [NCBI Gene 69601] {aka 2310011D08Rik, Aip1, mKIAA1743}, Smad1 (SMAD family member 1) [NCBI Gene 17125] {aka Mad1, Madh1, Madr1, Mlp1, MusMLP, dwf-A}, Rere (arginine glutamic acid dipeptide (RE) repeats) [NCBI Gene 68703] {aka 1110033A15Rik, ARG, ARP, ATN1L, Atr2, DNB1}, Trp53-ps (transformation related protein 53, pseudogene) [NCBI Gene 22060], MIR378-1 (microRNA mir-378-1) [NCBI Gene 100313042] {aka MIR378, bta-mir-378, bta-mir-378-1, mir-378-1}, Paip2 (polyadenylate-binding protein-interacting protein 2) [NCBI Gene 67869] {aka 2310050K10Rik}, GAPDH (glyceraldehyde-3-phosphate dehydrogenase) [NCBI Gene 2597] {aka G3PD, GAPD, HEL-S-162eP}, ZBTB7A (zinc finger and BTB domain containing 7A) [NCBI Gene 788491], MYH4 (myosin heavy chain 4) [NCBI Gene 408020], Casp2 (caspase 2) [NCBI Gene 12366] {aka CASP-2, ICH-1, NEDD-2, Nedd2}, Tgfb1 (transforming growth factor, beta 1) [NCBI Gene 21803] {aka TGF-beta1, TGFbeta1, Tgfb, Tgfb-1}, Aebp2 (AE binding protein 2) [NCBI Gene 11569] {aka B230313N05Rik}, Zbtb7a (zinc finger and BTB domain containing 7a) [NCBI Gene 16969] {aka 9030619K07Rik, 9130006G12Rik, FBI-1, Lrf, Pokemon, Zbtb7}, Pdk4 (pyruvate dehydrogenase kinase, isoenzyme 4) [NCBI Gene 27273], Tnf (tumor necrosis factor) [NCBI Gene 21926] {aka DIF, TNF-a, TNF-alpha, TNFSF2, TNFalpha, Tnfa}, Foxo1 (forkhead box O1) [NCBI Gene 56458] {aka Afxh, FKHR, Fkhr1, Foxo1a}, MMUT (methylmalonyl-CoA mutase) [NCBI Gene 4594] {aka MCM, MUT}, Myog (myogenin) [NCBI Gene 17928] {aka MYF4, bHLHc3, myo}, Mir182 (microRNA 182) [NCBI Gene 387177] {aka Mirn182, mir-182, mmu-mir-182}, MYF6 (myogenic factor 6) [NCBI Gene 281336] {aka MRF4}, Mir206 (microRNA 206) [NCBI Gene 387202] {aka Mirn206, mmu-mir-206}, MYOD1 (myogenic differentiation 1) [NCBI Gene 4654] {aka CMYO17, CMYP17, MYF3, MYOD, MYODRIF, PUM}, Myod1 (myogenic differentiation 1) [NCBI Gene 17927] {aka MYF3, MyoD, Myod-1, bHLHc1}, Nuak1 (NUAK family, SNF1-like kinase, 1) [NCBI Gene 77976] {aka B230104P22Rik, Omphk1}, MYH3 (myosin heavy chain 3) [NCBI Gene 281338], ACTE1 (actin epsilon 1) [NCBI Gene 528168], Arf4 (ARF GTPase 4) [NCBI Gene 11843], BMP4 (bone morphogenetic protein 4) [NCBI Gene 407216], MIR1825 (microRNA 1825) [NCBI Gene 100302183] {aka MIRN1825, hsa-mir-1825}, Dnaaf9 (dynein axonemal assembly factor 9) [NCBI Gene 228602] {aka 2810487F15Rik, 4930402H24Rik}, ZBTB7A (zinc finger and BTB domain containing 7A) [NCBI Gene 51341] {aka FBI-1, FBI1, LRF, MNDLFH, TIP21, ZBTB7}, Smad4 (SMAD family member 4) [NCBI Gene 17128] {aka D18Wsu70e, DPC4, Madh4}, MYF5 (myogenic factor 5) [NCBI Gene 281335], SESN2 (sestrin 2) [NCBI Gene 509863]
- **Diseases:** leiomyomatosis (MESH:D018231), cancer (MESH:D009369), prostate cancer (MESH:D011471)
- **Chemicals:** H (MESH:D006859), Giemsa (MESH:D001399), nitrogen (MESH:D009584), DAPI (MESH:C007293), CO2 (MESH:D002245), Palmitic acid (MESH:D019308), bicinchoninic acid (MESH:C047117), PBS (-), SDS (MESH:D012967), methanol (MESH:D000432), sugar (MESH:D000073893), glucose (MESH:D005947), TritonX-100 (MESH:D017830), Lipofectamine  2000 (MESH:C086724), water (MESH:D014867)
- **Species:** Mus musculus (house mouse, species) [taxon 10090], Bos taurus (bovine, species) [taxon 9913]
- **Cell lines:** 293 T — Homo sapiens (Human), Transformed cell line (CVCL_0063), CRL-1772 — Homo sapiens (Human), 5' 10' methylenetetrahydrofolate reductase deficiency, Finite cell line (CVCL_B3VW), C2C12 — Mus musculus (Mouse), Spontaneously immortalized cell line (CVCL_0188)

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## Figures

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## References

32 references — full list in the complete paper: https://tomesphere.com/paper/PMC12345297/full.md

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Source: https://tomesphere.com/paper/PMC12345297