Skip to main content
Download PDF
  • Research article
  • Open access
  • Published:

MicroRNA-877-5p promotes osteoblast differentiation by targeting EIF4G2 expression

Journal of Orthopaedic Surgery and Research volume 19, Article number: 134 (2024) Cite this article

Abstract

Stimulating bone formation potentially suggests therapeutics for orthopedic diseases including osteoporosis and osteoarthritis. Osteoblasts are key to bone remodeling because they act as the only bone-forming cells. miR-877-5p has a chondrocyte-improving function in osteoarthritis, but its effect on osteoblast differentiation is unknown. Here, miR-877-5p-mediated osteoblast differentiation was studied. Real-time reverse transcriptase-polymerase chain reaction was performed to measure miR-877-5p expression during the osteogenic differentiation of MC3T3-E1 cells. Osteoblast markers, including alkaline phosphatase (ALP), collagen type I a1 chain, and osteopontin, were measured and detected by alizarin red staining and ALP staining. Potential targets of miR-877-5p were predicted from three different algorithms: starBase (http://starbase.sysu.edu.cn/), PITA (http://genie.weizmann.ac.il/pubs/mir07/mir07_data.html), and miRanda (http://www.microrna.org/microrna/home.do). It was further verified by dual luciferase reporter gene assay. The experimental results found that miR-877-5p was upregulated during the osteogenic differentiation of MC3T3-E1 cells. Overexpression of miR-877-5p promoted osteogenic differentiation, which was characterized by increased cell mineralization, ALP activity, and osteogenesis-related gene expression. Knockdown of miR-877-5p produced the opposite result. Dual luciferase reporter gene assay showed that miR-877-5p directly targeted eukaryotic translation initiation factor 4γ2 (EIF4G2). Overexpression of EIF4G2 inhibited osteogenic differentiation and reversed the promoting effect of overexpression of miR-135-5p on osteogenic differentiation. These results indicate that miR-877-5p might have a therapeutic application related to its promotion of bone formation through targeting EIF4G2.

Introduction

Osteoporosis is a globally prevalent bone condition connected with bone resorption and loss of the bone microstructure, which can lead to bone fragility and increased bone fracture [1]. One of the key methods to manage the imbalance in bone mass is to stimulate osteoblast formation [2]. Osteoblasts, as the main cell type for bone formation, are the key to the metabolic homeostasis, growth, and injury repair of bone tissue [3,4,5], and some protein markers, such as alkaline phosphatase (ALP), collagen type I a1 chain (COL1A1), and osteopontin (OPN), are believed to have value as bone development biomarkers [6]. Therefore, investigation into the regulatory mechanisms behind osteoblast differentiation will aid in the diagnosis or treatment of osteoporosis.

Osteoblast differentiation is closely regulated by a variety of factors [7,8,9], among which miRNAs have recently received attention. miRNAs exert their regulatory functions by regulating their target genes at the post-transcriptional level, thereby participating in various biological processes [10,11,12,13,14]. miR-25 [15], miR-33-5p [16], miR-221, and miR-15b [17] have been revealed to act on corresponding targets, thus affecting the balance between bone formation and resorption, and therein regulating osteoblast differentiation. miR-877-5p is frequently downregulated in various cancers [18,19,20], and particularly, miR-877-5p has an alleviating impact on osteoarthritis chondrocytes [21]. In addition, miR-877-3p promotes osteoblast differentiation of MC3T3-E1 cells by targeting Smad7 [22]. However, the role of miR-877-3p in osteoblast differentiation has not been studied.

Translation of most mRNAs is regulated at the initiation level, a process that requires a protein complex called EIF4F [23]. Eukaryotic translation initiation factor 4γ (EIF4G) has two functional homologs in mammals, namely EIF4G1 and EIF4G2 [24]. EIF4G2 has been studied in osteoarthritic cartilage and has an association with chondrocytes in the pathogenesis of osteoarthritis [25]. Potential targets of miR-877-5p were predicted from three different algorithms: starBase (http://starbase.sysu.edu.cn/), PITA (http://genie.weizmann.ac.il/pubs/mir07/mir07_data.html), and miRanda (http://www.microrna.org/microrna/home.do). The results showed that miR-877-5p and EIF4G2 had targeted binding sites. Moreover, the targeting relationship between miR-877-5p and EIF4G2 was further verified by dual luciferase reporter gene assay.

Therefore, this study aimed to investigate the role of miR-877-5p and EIF4G2 in osteoblast differentiation and found that miR-877-5p promoted osteoblast differentiation through targeted regulation of EIF4G2 expression. Our results may be useful in enhancing new bone formation and designing treatments for pathological bone loss.

Methods

Cell culture

Mouse osteogenic MC3T3-E1 cells (Cell Bank of Chinese Academy of Sciences) were cultured in Dulbecco's modified Eagle medium containing 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 mg/ml streptomycin. Osteoblast differentiation induction medium was prepared by α-minimum essential medium (Hyclone), 10% FBS, 1% penicillin/streptomycin, 50 ug/mL l-ascorbic acid, 10 mM β-glycerophosphate, and 10 nM dexamethasone. At 3, 7, 14, and 21 d after osteogenic induction, miR-877-5p and EIF4G2 levels were checked.

MC3T3-E1 cell transfection

miR-877-5p mimic (5′-GUAGAGGAGAUGGCGCAGGG-3′), miR-877-5p inhibitor (5′-CCCUGCGCCAUCUCCUCUAC-3′), small interfering RNA (siRNA) targeting EIF4G2 (si-EIF4G2, sense, 5′-GCUUCUCGUUUCAGUGCUUTT-3′, antisense, 5′-AAGCACUGAAACGAGAAGCTT-3′), pcDNA 3.1 vector containing EIF4G2 (oe-EIF4G2), and their corresponding negative controls (GenePharma, Shanghai, China) were transfected into MC3T3-E1 cells at a concentration of 10 nM using Lipofectamine RNAiMAX transfection reagent (Invitrogen). After transfection, osteogenic induction was performed.

RNA quantification

Total RNA was extracted from cells using TRIzol reagent (Thermo fisher, USA). The purity and concentration of RNA were spectrophotometrically analyzed using a Nanodrop One (Thermo Fisher). To determine miRNA, cDNA was transcribed using the TaqMan MicroRNA Reverse Transcription Kit (Thermo Fisher). To determine mRNAs, cDNA was transcribed using the First Strand cDNA Synthesis Kit (Beyotime, China). PCR was performed on an ABI 7500 Real-Time PCR system (Thermo Fisher) using the TB Green Premix Ex Taq II (Taraka, Japan). U6 was the reference gene for miRNA in the cell, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was the reference gene for mRNA in the cell. The gene expressions were quantified using the 2−ΔΔCT method. Sangong (Shanghai, China) was commissioned to synthesize primers (Table 1).

Table 1 PCR primers
Full size table

Western blot

Total proteins were extracted using radioimmunoprecipitation assay buffer (pH 7.4) and separated on 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Proteins transferred to nitrocellulose membranes (Millipore) were mixed with primary antibodies EIF4G2 (1:1000, ab97302, Abcam) and GAPDH (1:1000, ab8245, Abcam) after being blocked with 5% low-fat milk. Next, horseradish peroxidase-conjugated secondary antibody (1:5000, ab205718, Abcam) was supplemented to develop protein bands using enhanced chemiluminescence reagents (Amersham Biosciences, Piscataway, NJ, USA).

ALP staining and alizarin red staining

For ALP staining, after induction for 14 days, cells were fixed with 95% ethanol and stained with BCIP/NBT solution according to the manufacturer's protocol (Beyotime Institute of Biotechnology) at room temperature for 2 h. The ALP-positive cells were stained blue/purple. Stained cells were visualized using the Canon IXUS210 camera (Canon, Inc.; magnification). In addition, an Alkaline Phosphatase Staining Kit (BestBio, Inc.) was used to detect the ALP activity, according to the manufacturer's protocol.

For alizarin red staining, after induction for 14 days, cells were washed one or two times with phosphate-buffered saline (PBS), fixed with 95% ethanol for 10 min, washed one or two times with PBS again, covered, and stained with 0.1% alizarin red solution for 10 min. Finally, they were rinsed with PBS and observed under an inverted light microscope. For quantification analysis, 10% hexadecyl pyridinium chloride monohydrate (CPC; Sigma-Aldrich; Merck KGaA) was used to dissolve the mineralized nodules and then the absorbance was measured at 540 nm using a Multiskan™ FC spectrophotometer (Thermo Fisher Scientific, Inc.).

Dual luciferase activity assay

The amplified EIF4G2 fragment containing the miR-877-5p binding site was cloned into the psi-CHEK2 vector (Promega) to obtain the EIF4G2 wild-type (WT). Also, an EIF4G2 mutant (MUT) vector was produced. MC3T3-E1 cells co-transfected with the above plasmids and miR-877-5p mimic or NC using Lipofectamine 2000 (Invitrogen). Luciferase activity was determined using a dual luciferase reporter assay system (Promega).

Statistical analysis

Statistical analysis was performed using PRISM v5.0 software (GraphPad, La Jolla, CA, USA). All data in three replicates are expressed as mean standard deviation and assessed by Wilcoxon test. P < 0.05 was considered statistically significant.

Results

High miR-877-5p and low EIF4G2 in osteoblast differentiation

At 3, 7, 14, and 21 d of osteoblast differentiation, miR-877-5p and EIF4G2 levels were quantified: miR-877-5p expression increased over time in MC3T3-E1 cells, whereas EIF4G2 expression decreased (Fig. 1A, B).

Fig. 1
figure 1

High miR-877-5p and low EIF4G2 during osteoblast differentiation. A/B: RT-qPCR or Western blot to determine miR-877-5p and EIF4G2 in MC3T3-E1 cells on days 3, 7, 14, and 21 of osteoblast differentiation; values are expressed as mean ± standard deviation; *P < 0.05 versus day 3

Full size image

Promoting effects of miR-877-5p on osteoblast differentiation

Transfection with miR-877-5p mimic or miR-877-5p inhibitor and their negative control (NC) was implemented before osteoblast differentiation. RT-qPCR results confirmed that the vectors were successfully transfected (Fig. 2A). After 14 days of osteogenic induction, alizarin red staining showed that mineralization of MC3T3-E1 cells increased after miR-877-5p upregulation (Fig. 2B, C). After 14 days of osteogenic induction, ALP staining was performed. The results showed that the activity of ALP increased after upregulation of miR-877-5p (Fig. 2D, E). In addition, miR-877-5p upregulation elevated mRNA expression of osteoblast markers ALP, COLIA1, OPN, β-catenin, and RUNX2 (Fig. 2F–H, Additional file 1: Fig. S1A). In contrast to that, miR-877-5p downregulation caused the negative results (Fig. 2B–H, Supplementary Fig. 1A).

Fig. 2
figure 2

Promoting effects of miR-877-5p on osteoblast differentiation. (A) RT-qPCR to verify the successful transfection of miR-877-5p mimic or inhibitor; (B/C) alizarin red staining to detect the mineralization of MC3T3-E1 cells; (D) ALP activity assay; (E–G) RT-qPCR detection of ALP, COLIA1, and OPN; values are expressed as mean ± standard deviation; *P < 0.05 versus mimic NC; #P < 0.05 versus inhibitor NC

Full size image

A negative relation between miR-877-5p and EIF4G2

Potential targets of miR-877-5p were predicted from three different algorithms: starBase (http://starbase.sysu.edu.cn/), PITA (http://genie.weizmann.ac.il/pubs/mir07/mir07_data.html), and miRanda (http://www.microrna.org/microrna/home.do). A putative miR-877-5p binding site was found in the EIF4G2 3’UTR (Fig. 3A). Luciferase activity was reduced in MC3T3-E1 cells co-transfected with miR-877-5p mimic and EIF4G2-WT (Fig. 3B). In addition, EIF4G2 expression was suppressed after miR-877-5p mimic transfection in MC3T3-E1 cells, while those transfected with miR-877-5p inhibitor had elevated EIF4G2 expression (Fig. 3C).

Fig. 3
figure 3

A negative relation between miR-877-5p and EIF4G2. A: starBase predicted the binding sites of miR-877-5p and EIF4G2; B: dual luciferase reporter gene detection confirmed that miR-877-5p targeted binding to EIF4G2; C: RT-qPCR and Western blot detection of EIF4G2; the values are expressed as mean ± standard deviation; *P < 0.05 versus mimic NC; #P < 0.05 versus inhibitor NC

Full size image

Suppressive effects of EIF4G2 on osteoblast differentiation

Transfection with si-EIF4G2 or oe-EIF4G2 was performed before osteoblast differentiation induction, and the vectors were successfully transfected (Fig. 4A). Responded to EIF4G2 knockdown, increased mineralization, ALP activity, and expression of osteoblast markers were seen in MC3T3-E1 cells; EIF4G2 overexpression had the opposite effect (Fig. 4B-H, Additional file 1: Fig. S1B).

Fig. 4
figure 4

Suppressive effects of EIF4G2 on osteoblast differentiation. (A) RT-qPCR and Western blot to verify the successful transfection of si-EIF4G2 or oe-EIF4G2; (B/C) alizarin red staining to detect the mineralization of MC3T3-E1 cells; (D) ALP activity assay; (E–G) RT-qPCR detection of ALP, COLIA1, and OPN; the values are expressed as mean ± standard deviation; *P < 0.05 versus si-NC; #P < 0.05 versus oe-NC

Full size image

Overexpression of EIF4G2 reverses the promotion of miR-877-5p upregulation on osteoblast differentiation

To investigate the relationship between miR-877-5p and EIF4G2 in more detail, co-transfection with miR-877-5p mimic and oe-EIF4G2 was carried out in MC3T3-E1 cells. oe-EIF4G2 not only abolished the suppressive impact of miR-877-5p mimic on EIF4G2 expression (Fig. 5A), but the promoting effect on osteoblast differentiation (Fig. 5B-G, Additional file 1: Fig. S1C).

Fig. 5
figure 5

EIF4G2 reverses the promotion of miR-877-5p upregulation on osteoblast differentiation. (A) RT-qPCR and Western blot to verify the successful co-transfection of miR-877-5p mimic and oe-EIF4G2; (B/C) alizarin red staining to detect the mineralization of MC3T3-E1 cells; D: ALP activity assay; (E–G) RT-qPCR detection of ALP, COLIA1, and OPN; the values are expressed as mean ± standard deviation; *P < 0.05 versus miR-877-5p mimic + oe-NC

Full size image

Discussion

Osteoblast differentiation is an essential component of bone remodeling to maintain bone health [26]. Inappropriate regulation of bone formation is associated with diseases such as osteoporosis, osteoarthritis, and bone cancer. The osteoblast differentiation process involves the regulation of gene expression, osteoblast differentiation marker genes, and mineral deposition [7,8,9]. Currently, many miRNAs emphasize great roles in skeletal development and homeostasis, especially in osteoblast differentiation [27,28,29]. Therefore, our study investigated miR-877-5p in osteoblast differentiation from the perspective of gene regulation and concluded that miR-877-5p promotes osteoblast differentiation by negatively regulating EIF4G2 expression. This is the first demonstration of the role and potential mechanism of miR-877-5p in osteogenesis.

MC3T3-E1 osteoblast cell line can be used as a host for in vitro studies of bone remodeling and formation [30,31,32,33] due to its pre-osteoblast phenotype, and its subclone 14 can mineralize the collagen extracellular matrix [34]. Therefore, mouse osteogenic MC3T3-E1 cells were applied to induce osteoblast differentiation model. Numerous studies have shown that miRNAs could act as key modulators in osteoblastic differentiation. MiR-497-5p stimulates osteoblast differentiation through HMGA2-mediated JNK pathway [30]. MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells [35]. In addition, miR-532-3p inhibits osteogenic differentiation in MC3T3-E1 cells by downregulating ETS1 [36]. In this study, it was found that miR-877-5p was upregulated during the osteogenic differentiation of MC3T3-E1. Osteogenesis-related markers in osteoblast differentiation, including OPN, COLIA1, and ALP, are at high levels during extracellular matrix maturation and mineralization [37, 38]. Furthermore, mineralization of the extracellular matrix represents the end stage of osteoblast differentiation and is considered a sign of osteoblast maturation [39, 40]. Here, we found that miR-877-5p upregulation increased the mineralization of MC3T3-E1 cells and induced ALP activity and osteoblast markers’ expression.

miRNAs reduce the translation and/or degradation of target gene expression by targeting the 3′-UTR of mRNA [41]. To explore the molecular mechanism of miR-877-5p regulating osteogenic differentiation of MC3T3-E1 cells, potential targets of miR-877-5p were predicted from three different algorithms: starBase (http://starbase.sysu.edu.cn/), PITA (http://genie.weizmann.ac.il/pubs/mir07/mir07_data.html), and miRanda (http://www.microrna.org/microrna/home.do). EIF4G2, a downregulated actor during osteogenic differentiation, was validated as a downstream gene of miR-877-5p. EIF4G2 plays an important role during cell mitosis [42] and can be involved in different types of cell differentiation, such as myeloid monocytes [43], myogenic cells [44], and embryonic stem cells [45]. Recently, Hu et al. found that EIF4G2 has regulatory effects on chondrocyte viability, colony formation, and migration [46]. For novelty, our results demonstrated that downregulation of EIF4G2 promoted osteoblast differentiation, whereas upregulation of EIF4G2 inhibited osteoblast differentiation, and upregulation of EIF4G2 reversed the promoting effect of upregulation of miR-877-5p on osteoblast differentiation.

Nonetheless, the results are only applicable to in vitro osteoblast models and it is unclear whether miR-877-5p and EIF4G2 have similar effects in animal models. Since bone formation can be mediated by the recruitment of mesenchymal stem cells [47], the process of MSC differentiation into osteoblasts should be further explored.

Conclusion

We reveal for the novelty that miR-877-5p promotes osteoblast differentiation by targeting and negatively regulating EIF4G2 expression. Therefore, therapeutic strategies targeting miR-877-5p may promote bone formation and may be effective in treating orthopedic diseases.

Availability of data and materials

Data are available from the corresponding author on request.

References

  1. Oheim R, Schinke T, Amling M, Pogoda P. Can we induce osteoporosis in animals comparable to the human situation? Injury. 2016;47(Suppl 1):S3-9.

    Article  PubMed  Google Scholar 

  2. Oh JH, Ahn BN, Karadeniz F, Kim JA, Lee JI, Seo Y, et al. Phlorofucofuroeckol a from edible brown alga ecklonia cava enhances osteoblastogenesis in bone marrow-derived human mesenchymal stem cells. Mar Drugs. 2019;17(10):1.

    Article  Google Scholar 

  3. Fang S, Deng Y, Gu P, Fan X. MicroRNAs regulate bone development and regeneration. Int J Mol Sci. 2015;16(4):8227–53.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. Ruan C, Hu N, Ma Y, Li Y, Liu J, Zhang X, et al. The interfacial pH of acidic degradable polymeric biomaterials and its effects on osteoblast behavior. Sci Rep. 2017;7(1):6794.

    Article  ADS  PubMed  PubMed Central  Google Scholar 

  5. Jiang Z, Wang H, Qi G, Jiang C, Chen K, Yan Z. Iron overload-induced ferroptosis of osteoblasts inhibits osteogenesis and promotes osteoporosis: an in vitro and in vivo study. IUBMB Life. 2022;74(11):1052–69.

    Article  PubMed  CAS  Google Scholar 

  6. Shimoda A, Sawada SI, Sasaki Y, Akiyoshi K. Exosome surface glycans reflect osteogenic differentiation of mesenchymal stem cells: profiling by an evanescent field fluorescence-assisted lectin array system. Sci Rep. 2019;9(1):11497.

    Article  ADS  PubMed  PubMed Central  Google Scholar 

  7. Rutkovskiy A, Stensløkken KO, Vaage IJ. Osteoblast differentiation at a glance. Med Sci Monit Basic Res. 2016;22:95–106.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Komori T. Regulation of osteoblast differentiation by transcription factors. J Cell Biochem. 2006;99(5):1233–9.

    Article  PubMed  CAS  Google Scholar 

  9. Marie PJ. Transcription factors controlling osteoblastogenesis. Arch Biochem Biophys. 2008;473(2):98–105.

    Article  PubMed  CAS  Google Scholar 

  10. Jin T, Lu Y, He QX, Wang H, Li BF, Zhu LY, et al. The role of MicroRNA, miR-24, and Its target CHI3L1 in osteomyelitis caused by Staphylococcus aureus. J Cell Biochem. 2015;116(12):2804–13.

    Article  PubMed  CAS  Google Scholar 

  11. Giordano L, Porta GD, Peretti GM, Maffulli N. Therapeutic potential of microRNA in tendon injuries. Br Med Bull. 2020;133(1):79–94.

    Article  PubMed  CAS  Google Scholar 

  12. Oliviero A, Della Porta G, Peretti GM, Maffulli N. MicroRNA in osteoarthritis: physiopathology, diagnosis and therapeutic challenge. Br Med Bull. 2019;130(1):137–47.

    Article  PubMed  CAS  Google Scholar 

  13. Gargano G, Oliviero A, Oliva F, Maffulli N. Small interfering RNAs in tendon homeostasis. Br Med Bull. 2021;138(1):58–67.

    Article  PubMed  CAS  Google Scholar 

  14. Gargano G, Oliva F, Oliviero A, Maffulli N. Small interfering RNAs in the management of human rheumatoid arthritis. Br Med Bull. 2022;142(1):34–43.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Li X, Ji J, Wei W, Liu L. MiR-25 promotes proliferation, differentiation and migration of osteoblasts by up-regulating Rac1 expression. Biomed Pharmacother. 2018;99:622–8.

    Article  PubMed  CAS  Google Scholar 

  16. Wang H, Sun Z, Wang Y, Hu Z, Zhou H, Zhang L, et al. miR-33-5p, a novel mechano-sensitive microRNA promotes osteoblast differentiation by targeting Hmga2. Sci Rep. 2016;6:23170.

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  17. Lu X, Zhang Y, Zheng Y, Chen B. The miRNA-15b/USP7/KDM6B axis engages in the initiation of osteoporosis by modulating osteoblast differentiation and autophagy. J Cell Mol Med. 2021;25(4):2069–81.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  18. Ma J, Li Q, Li Y. CircRNA PRH1-PRR4 stimulates RAB3D to regulate the malignant progression of NSCLC by sponging miR-877-5p. Thorac Cancer. 2022;13(5):690–701.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Yang J, Tian S, Wang B, Wang J, Cao L, Wang Q, et al. CircPIK3C2A facilitates the progression of glioblastoma via targeting miR-877-5p/FOXM1 axis. Front Oncol. 2021;11: 801776.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  20. Wu T, Sun Y, Sun Z, Li S, Wang W, Yu B, et al. Hsa_circ_0042823 accelerates cancer progression via miR-877-5p/FOXM1 axis in laryngeal squamous cell carcinoma. Ann Med. 2021;53(1):960–70.

    Article  PubMed  Google Scholar 

  21. Zhu S, Deng Y, Gao H, Huang K, Nie Z. miR-877-5p alleviates chondrocyte dysfunction in osteoarthritis models via repressing FOXM1. J Gene Med. 2020;22(11): e3246.

    Article  PubMed  CAS  Google Scholar 

  22. He G, Chen J, Huang D. miR-877-3p promotes TGF-β1-induced osteoblast differentiation of MC3T3-E1 cells by targeting Smad7. Exp Ther Med. 2019;18(1):312–9.

    PubMed  PubMed Central  CAS  Google Scholar 

  23. Wang Z, Ding X, Cao F, Zhang X, Wu J. Bone mesenchymal stem cells promote extracellular matrix remodeling of degenerated nucleus pulposus cells via the miR-101-3p/EIF4G2 axis. Front Bioeng Biotechnol. 2021;9: 642502.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Caron S, Charon M, Cramer E, Sonenberg N, Dusanter-Fourt I. Selective modification of eukaryotic initiation factor 4F (eIF4F) at the onset of cell differentiation: recruitment of eIF4GII and long-lasting phosphorylation of eIF4E. Mol Cell Biol. 2004;24(11):4920–8.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Gao S, Liu L, Zhu S, Wang D, Wu Q, Ning G, et al. MicroRNA-197 regulates chondrocyte proliferation, migration, and inflammation in pathogenesis of osteoarthritis by targeting EIF4G2. Biosci Rep. 2020;40(9):1.

    Article  Google Scholar 

  26. Narayanan A, Srinaath N, Rohini M, Selvamurugan N. Regulation of Runx2 by MicroRNAs in osteoblast differentiation. Life Sci. 2019;232: 116676.

    Article  PubMed  CAS  Google Scholar 

  27. Gámez B, Rodriguez-Carballo E, Ventura F. MicroRNAs and post-transcriptional regulation of skeletal development. J Mol Endocrinol. 2014;52(3):R179–97.

    Article  PubMed  Google Scholar 

  28. An F, Wang X, Wang C, Liu Y, Sun B, Zhang J, et al. Research progress on the role of lncRNA-miRNA networks in regulating adipogenic and osteogenic differentiation of bone marrow mesenchymal stem cells in osteoporosis. Front Endocrinol (Lausanne). 2023;14:1210627.

    Article  PubMed  Google Scholar 

  29. Zhang Q, Long Y, Jin L, Li C, Long J. Non-coding RNAs regulate the BMP/Smad pathway during osteogenic differentiation of stem cells. Acta Histochem. 2023;125(1): 151998.

    Article  PubMed  CAS  Google Scholar 

  30. Zhao H, Yang Y, Wang Y, Feng X, Deng A, Ou Z, et al. MicroRNA-497-5p stimulates osteoblast differentiation through HMGA2-mediated JNK signaling pathway. J Orthop Surg Res. 2020;15(1):515.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Czekanska EM, Stoddart MJ, Richards RG, Hayes JS. In search of an osteoblast cell model for in vitro research. Eur Cell Mater. 2012;24:1–17.

    Article  PubMed  CAS  Google Scholar 

  32. Long Z, Dou P, Cai W, Mao M, Wu R. MiR-181a-5p promotes osteogenesis by targeting BMP3. Aging (Albany NY). 2023;15(3):734–47.

    Article  PubMed  CAS  Google Scholar 

  33. Ma S, Li S, Zhang Y, Nie J, Cao J, Li A, et al. BMSC-derived exosomal CircHIPK3 promotes osteogenic differentiation of MC3T3-E1 cells via mitophagy. Int J Mol Sci. 2023;24(3):1.

    Article  Google Scholar 

  34. Casati L, Pagani F, Maggi R, Ferrucci F, Sibilia V. Food for bone: evidence for a role for delta-tocotrienol in the physiological control of osteoblast migration. Int J Mol Sci. 2020;21(13):1.

    Article  Google Scholar 

  35. Yin N, Zhu L, Ding L, Yuan J, Du L, Pan M, et al. MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells. Cell Mol Biol Lett. 2019;24:51.

    Article  ADS  PubMed  PubMed Central  Google Scholar 

  36. Fan Q, Li Y, Sun Q, Jia Y, He C, Sun T. miR-532-3p inhibits osteogenic differentiation in MC3T3-E1 cells by downregulating ETS1. Biochem Biophys Res Commun. 2020;525(2):498–504.

    Article  PubMed  CAS  Google Scholar 

  37. Padiolleau L, Chanseau C, Durrieu S, Chevallier P, Laroche G, Durrieu MC. Single or mixed tethered peptides to promote hMSC differentiation toward osteoblastic lineage. ACS Appl Bio Mater. 2018;1(6):1800–9.

    Article  PubMed  CAS  Google Scholar 

  38. Hwang JH, Park YS, Kim HS, Kim DH, Lee SH, Lee CH, et al. Yam-derived exosome-like nanovesicles stimulate osteoblast formation and prevent osteoporosis in mice. J Control Release. 2023;355:184–98.

    Article  PubMed  CAS  Google Scholar 

  39. Quarles LD, Yohay DA, Lever LW, Caton R, Wenstrup RJ. Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: an in vitro model of osteoblast development. J Bone Miner Res. 1992;7(6):683–92.

    Article  PubMed  CAS  Google Scholar 

  40. Li Y, Su J, Sun W, Cai L, Deng Z. AMP-activated protein kinase stimulates osteoblast differentiation and mineralization through autophagy induction. Int J Mol Med. 2018;41(5):2535–44.

    PubMed  PubMed Central  CAS  Google Scholar 

  41. Bartel DP. MicroRNAs: target recognition and regulatory functions. Cell. 2009;136(2):215–33.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  42. Coldwell MJ, Cowan JL, Vlasak M, Mead A, Willett M, Perry LS, et al. Phosphorylation of eIF4GII and 4E-BP1 in response to nocodazole treatment: a reappraisal of translation initiation during mitosis. Cell Cycle. 2013;12(23):3615–28.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  43. Emmrich S, Engeland F, El-Khatib M, Henke K, Obulkasim A, Schöning J, et al. miR-139-5p controls translation in myeloid leukemia through EIF4G2. Oncogene. 2016;35(14):1822–31.

    Article  PubMed  CAS  Google Scholar 

  44. Sanson M, Vu Hong A, Massourides E, Bourg N, Suel L, Amor F, et al. miR-379 links glucocorticoid treatment with mitochondrial response in Duchenne muscular dystrophy. Sci Rep. 2020;10(1):9139.

    Article  ADS  PubMed  PubMed Central  CAS  Google Scholar 

  45. Yoffe Y, David M, Kalaora R, Povodovski L, Friedlander G, Feldmesser E, et al. Cap-independent translation by DAP5 controls cell fate decisions in human embryonic stem cells. Genes Dev. 2016;30(17):1991–2004.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  46. Hu W, Zhang W, Li F, Guo F, Chen A. miR-139 is up-regulated in osteoarthritis and inhibits chondrocyte proliferation and migration possibly via suppressing EIF4G2 and IGF1R. Biochem Biophys Res Commun. 2016;474(2):296–302.

    Article  PubMed  CAS  Google Scholar 

  47. Bartold M, Gronthos S, Haynes D, Ivanovski S. Mesenchymal stem cells and biologic factors leading to bone formation. J Clin Periodontol. 2019;46(Suppl 21):12–32.

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

Not applicable.

Funding

In 2021, a project was funded by Changshu Science and Technology Bureau, Research on the mechanism of Circ_FKBP5 regulating miR-205-5p/RUNX2 axis to promote osteogenic differentiation of BMSC cells, No. 202131, and Changshu Municipal Health Commission Science and Technology Program, MK-3903 based on circBRAF-AMPK pathway to interfere with osteoblast injury in femoral head necrosis, ID: csws202004. In 2021, the Natural Science Foundation of Nanjing University of Traditional Chinese Medicine was approved. The abnormality of circBRAF-AMPK pathway is involved in the mechanism of dexamethasone-induced osteoblast injury. No. XZR2020064. Topic of Suzhou Municipal Health Commission, Key Clinical Diagnosis and Treatment Project, No. LCZX201923, Treatment of Osteoporosis Vertebral Fractures with "Eggshell" Technology Optimized.

Author information

Author notes

  1. YingChao Shen, Yang Zhang, and Qiang Wang have contributed equally to this work.

Authors and Affiliations

  1. Department of Orthopaedics, Changshu Hospital Affiliated to Nanjing University of Chinese Medicine, No. 6 Huanghe Road, ChangShu City, 215500, China

    YingChao Shen, Qiang Wang, XiaoWei Jiang & Bin Luo

  2. School of Biology and Food Engineering, Changshu Institute of Technology, Changshu City, 215500, Jiangsu, China

    Yang Zhang

  3. Department of Hand and Foot Surgery, The Second Affiliated Hospital of Soochow University, Jiangsu Province, No. 1055 Sanxiang Road, Suzhou City, 215004, China

    Bo Jiang

Authors
  1. YingChao Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qiang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Bo Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. XiaoWei Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bin Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

YCS, YZ, and QW designed the research study and wrote the manuscript. BL, BJ, and XW Jiang performed the research. QW, BJ, and XWJ provided help and advice. YCS, YZ, and BL analyzed the data. All authors contributed to editorial changes in the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Bo Jiang or XiaoWei Jiang.

Ethics declarations

Ethical approval and consent to participate

Not applicable.

Competing interests

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Additional file 1: Supplementary Fig. S1

. Regulatory effects of miR-877-5p/EIF4G2 axis on β-catenin and RUNX2 expression. A-C: RT-qPCR to detect β-catenin and RUNX2; the values are expressed as mean ± standard deviation; *P < 0.05 versus mimic NC; #P < 0.05 versus inhibitor NC; + P < 0.05 versus si-NC; ^P < 0.05 versus oe-NC; & P < 0.05 versus miR-877-5p mimic + oe-NC.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shen, Y., Zhang, Y., Wang, Q. et al. MicroRNA-877-5p promotes osteoblast differentiation by targeting EIF4G2 expression. J Orthop Surg Res 19, 134 (2024). https://doi.org/10.1186/s13018-023-04396-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s13018-023-04396-y

Keywords

Journal of Orthopaedic Surgery and Research

ISSN: 1749-799X