MicroRNA-200c-5p Regulates Migration and Differentiation of Myoblasts via Targeting Adamts5 in Skeletal Muscle Regeneration and Myogenesis
Abstract
:1. Introduction
2. Results
2.1. Identifying miR-200c-5p as a Candidate Regulator for Skeletal Muscle Regeneration
2.2. miR-200c-5p Regulates the Migration of C2C12 Myoblast, but Does Not Affect Its Proliferation
2.3. miR-200c-5p Modulates the Differentiation of C2C12 Myoblast
2.4. Adamts5 Is a Direct Target for miR-200c-5p
2.5. Adamts5 Regulates the Migration of C2C12 Myoblast but Does Not Affect Its Proliferation
2.6. Adamts5 Is Involved in the Differentiation of C2C12 Myoblast
2.7. miR-200c-5p Regulates the Migration and Differentiation of C2C12 Myoblast via Targeting Adamts5
3. Discussion
4. Materials and Methods
4.1. Study Design
4.2. Participants
4.3. Study Variables
4.3.1. Muscle Injury Model of Mice
4.3.2. Hematoxylin-Eosin (H&E) Staining
4.3.3. Cell Culture and Transfection
4.3.4. Plasmid and Oligonucleotides
4.3.5. RNA Extraction and Real-Time Quantitative PCR (qPCR)
4.3.6. Protein Extraction and Western Blot
4.3.7. Wound Healing Assay
4.3.8. Trans well
4.3.9. Cell Counting Kit-8 Proliferation Assay
4.3.10. Immunofluorescence Assay
4.3.11. RNA Immunoprecipitation (RIP)
4.3.12. Double Luciferase Assay
4.3.13. Online Prediction of Target Genes
4.4. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Frontera, W.R.; Ochala, J. Skeletal muscle: A brief review of structure and function. Calcif. Tissue Int. 2015, 96, 183–195. [Google Scholar] [CrossRef] [PubMed]
- Exeter, D.; Connell, D.A. Skeletal muscle: Functional anatomy and pathophysiology. Semin. Musculoskelet. Radiol. 2010, 14, 97–105. [Google Scholar] [CrossRef] [PubMed]
- Clarkson, P.M.; Hubal, M.J. Exercise-induced muscle damage in humans. Am. J. Phys. Med. Rehabil. 2002, 81, S52–S69. [Google Scholar] [CrossRef] [PubMed]
- Baghdadi, M.B.; Tajbakhsh, S. Regulation and phylogeny of skeletal muscle regeneration. Dev. Biol. 2018, 433, 200–209. [Google Scholar] [CrossRef]
- Turner, N.J.; Badylak, S.F. Regeneration of skeletal muscle. Cell Tissue Res. 2012, 347, 759–774. [Google Scholar] [CrossRef]
- Zhao, Y.; Chen, M.; Lian, D.; Li, Y.; Li, Y.; Wang, J.; Deng, S.; Yu, K.; Lian, Z. Non-Coding RNA Regulates the Myogenesis of Skeletal Muscle Satellite Cells, Injury Repair and Diseases. Cells 2019, 8, 988. [Google Scholar] [CrossRef] [Green Version]
- Goncalves, T.; Armand, A.S. Non-coding RNAs in skeletal muscle regeneration. Noncoding RNA Res. 2017, 2, 56–67. [Google Scholar] [CrossRef]
- Yu, X.; Zuo, Q. MicroRNAs in the regeneration of skeletal muscle. Front. Biosci. Landmark 2013, 18, 608–615. [Google Scholar] [CrossRef] [Green Version]
- Aranega, A.E.; Lozano-Velasco, E.; Rodriguez-Outeirino, L.; Ramirez, D.A.F.; Franco, D.; Hernandez-Torres, F. MiRNAs and Muscle Regeneration: Therapeutic Targets in Duchenne Muscular Dystrophy. Int. J. Mol. Sci. 2021, 22, 4236. [Google Scholar] [CrossRef]
- Lv, W.; Jin, J.; Xu, Z.; Luo, H.; Guo, Y.; Wang, X.; Wang, S.; Zhang, J.; Zuo, H.; Bai, W.; et al. lncMGPF is a novel positive regulator of muscle growth and regeneration. J. Cachexia Sarcopenia Muscle 2020, 11, 1723–1746. [Google Scholar] [CrossRef]
- Tajbakhsh, S. lncRNA-Encoded Polypeptide SPAR(s) with mTORC1 to Regulate Skeletal Muscle Regeneration. Cell Stem Cell 2017, 20, 428–430. [Google Scholar] [CrossRef] [Green Version]
- Yan, J.Y.; Yang, Y.L.; Fan, X.H.; Liang, G.M.; Wang, Z.S.; Li, J.J.; Wang, L.Y.; Chen, Y.; Adetula, A.A.; Tang, Y.J.; et al. circRNAome profiling reveals circFgfr2 regulates myogenesis and muscle regeneration via a feedback loop. J. Cachexia Sarcopenia Muscle 2022, 13, 696–712. [Google Scholar] [CrossRef] [PubMed]
- Yan, J.Y.; Yang, Y.L.; Fan, X.H.; Tang, Y.J.; Tang, Z.L. Sp1-Mediated circRNA circHipk2 Regulates Myogenesis by Targeting Ribosomal Protein Rpl7. Genes 2021, 12, 696. [Google Scholar] [CrossRef] [PubMed]
- Zhang, Z.; Fan, Y.X.; Deng, K.P.; Liang, Y.X.; Zhang, G.M.; Gao, X.X.; El-Samahy, M.A.; Zhang, Y.L.; Deng, M.T.; Wang, F. Circular RNA circUSP13 sponges miR-29c to promote differentiation and inhibit apoptosis of goat myoblasts by targeting IGF1. FASEB J. 2022, 36, e22097. [Google Scholar] [CrossRef] [PubMed]
- Tan, Y.; Shen, L.Y.; Gan, M.L.; Fan, Y.; Cheng, X.; Zheng, T.; Niu, L.L.; Chen, L.; Jiang, D.M.; Li, X.W.; et al. Downregulated miR-204 Promotes Skeletal Muscle Regeneration. Biomed Res. Int. 2020, 2020, 3183296. [Google Scholar] [CrossRef] [PubMed]
- Cheng, N.; Liu, C.; Li, Y.; Gao, S.; Han, Y.C.; Wang, X.; Du, J.; Zhang, C. MicroRNA-223-3p promotes skeletal muscle regeneration by regulating inflammation in mice. J. Biol. Chem. 2020, 295, 10212–10223. [Google Scholar] [CrossRef] [PubMed]
- Nie, M.; Liu, J.; Yang, Q.; Seok, H.Y.; Hu, X.; Deng, Z.L.; Wang, D.Z. MicroRNA-155 facilitates skeletal muscle regeneration by balancing pro- and anti-inflammatory macrophages. Cell Death Dis. 2016, 7, e2261. [Google Scholar] [CrossRef] [Green Version]
- Zhang, Y.; Yao, Y.; Wang, Z.; Lu, D.; Zhang, Y.; Adetula, A.A.; Liu, S.; Zhu, M.; Yang, Y.; Fan, X.; et al. MiR-743a-5p regulates differentiation of myoblast by targeting Mob1b in skeletal muscle development and regeneration. Genes Dis. 2022, 9, 1038–1048. [Google Scholar] [CrossRef]
- Zhang, Y.; Wang, Y. The correlation of plasma microRNA-200 family expressions with risk and disease severity of lupus nephritis. Eur. Rev. Med. Pharmacol. Sci. 2021, 25, 3641. [Google Scholar]
- Chen, G.; Zhang, W.; Ben, Y. Identification of Key Regulators of Hepatitis C Virus-Induced Hepatocellular Carcinoma by Integrating Whole-Genome and Transcriptome Sequencing Data. Front. Genet. 2021, 12, 741608. [Google Scholar] [CrossRef]
- Li, Y.; Bai, W.; Zhang, J. MiR-200c-5p suppresses proliferation and metastasis of human hepatocellular carcinoma (HCC) via suppressing MAD2L1. Biomed. Pharmacother. 2017, 92, 1038–1044. [Google Scholar] [CrossRef]
- Liu, Y.; Wang, J.; Zhou, X.; Cao, H.; Zhang, X.; Huang, K.; Li, X.; Yang, G.; Shi, X. miR-324-5p Inhibits C2C12 cell Differentiation and Promotes Intramuscular Lipid Deposition through lncDUM and PM20D1. Mol. Ther. Nucleic. Acids 2020, 22, 722–732. [Google Scholar] [CrossRef]
- Cai, B.; Ma, M.; Zhou, Z.; Kong, S.; Zhang, J.; Zhang, X.; Nie, Q. circPTPN4 regulates myogenesis via the miR-499-3p/NAMPT axis. J. Anim. Sci. Biotechnol. 2022, 13, 2. [Google Scholar] [CrossRef]
- Tidball, J.G. Mechanisms of muscle injury, repair, and regeneration. Compr. Physiol. 2011, 1, 2029–2062. [Google Scholar]
- Weskamp, K.; Olwin, B.B.; Parker, R. Post-Transcriptional Regulation in Skeletal Muscle Development, Repair, and Disease. Trends Mol. Med. 2021, 27, 469–481. [Google Scholar] [CrossRef]
- Stupka, N.; Kintakas, C.; White, J.D.; Fraser, F.W.; Hanciu, M.; Aramaki-Hattori, N.; Martin, S.; Coles, C.; Collier, F.; Ward, A.C.; et al. Versican processing by a disintegrin-like and metalloproteinase domain with thrombospondin-1 repeats proteinases-5 and -15 facilitates myoblast fusion. J. Biol. Chem. 2013, 288, 1907–1917. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mukund, K.; Subramaniam, S. Skeletal muscle: A review of molecular structure and function, in health and disease. Wiley Interdiscip. Rev. Syst. Biol. Med. 2020, 12, e1462. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mok, G.F.; Lozano-Velasco, E.; Munsterberg, A. microRNAs in skeletal muscle development. Semin. Cell Dev. Biol. 2017, 72, 67–76. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Shibasaki, H.; Imamura, M.; Arima, S.; Tanihata, J.; Kuraoka, M.; Matsuzaka, Y.; Uchiumi, F.; Tanuma, S.I.; Takeda, S. Characterization of a novel microRNA, miR-188, elevated in serum of muscular dystrophy dog model. PLoS ONE 2019, 14, e211597. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chen, J.F.; Tao, Y.; Li, J.; Deng, Z.; Yan, Z.; Xiao, X.; Wang, D.Z. microRNA-1 and microRNA-206 regulate skeletal muscle satellite cell proliferation and differentiation by repressing Pax7. J. Cell Biol. 2010, 190, 867–879. [Google Scholar] [CrossRef] [Green Version]
- Boettger, T.; Wust, S.; Nolte, H.; Braun, T. The miR-206/133b cluster is dispensable for development, survival and regeneration of skeletal muscle. Skelet. Muscle 2014, 4, 23. [Google Scholar] [CrossRef] [Green Version]
- Ma, G.; Wang, Y.; Li, Y.; Cui, L.; Zhao, Y.; Zhao, B.; Li, K. MiR-206, a key modulator of skeletal muscle development and disease. Int. J. Biol. Sci. 2015, 11, 345–352. [Google Scholar] [CrossRef]
- Dey, P.; Soyer, M.A.; Dey, B.K. MicroRNA-24-3p promotes skeletal muscle differentiation and regeneration by regulating HMGA1. Cell. Mol. Life Sci. 2022, 79, 170. [Google Scholar] [CrossRef]
- Dey, B.K.; Gagan, J.; Yan, Z.; Dutta, A. miR-26a is required for skeletal muscle differentiation and regeneration in mice. Genes Dev. 2012, 26, 2180–2191. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhai, L.; Wu, R.; Han, W.; Zhang, Y.; Zhu, D. miR-127 enhances myogenic cell differentiation by targeting S1PR3. Cell Death Dis. 2017, 8, e2707. [Google Scholar] [CrossRef] [Green Version]
- Kong, D.; He, M.; Yang, L.; Zhou, R.; Yan, Y.Q.; Liang, Y.; Teng, C.B. MiR-17 and miR-19 cooperatively promote skeletal muscle cell differentiation. Cell. Mol. Life Sci. 2019, 76, 5041–5054. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- D’Agostino, M.; Torcinaro, A.; Madaro, L.; Marchetti, L.; Sileno, S.; Beji, S.; Salis, C.; Proietti, D.; Imeneo, G.; Capogrossi, C.M.; et al. Role of miR-200c in Myogenic Differentiation Impairment via p66Shc: Implication in Skeletal Muscle Regeneration of Dystrophic mdx Mice. Oxid. Med. Cell. Longev. 2018, 2018, 4814696. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lu, M.Y.; Yu, C.C.; Chen, P.Y.; Hsieh, P.L.; Peng, C.Y.; Liao, Y.W.; Yu, C.H.; Lin, K.H. miR-200c inhibits the arecoline-associated myofibroblastic transdifferentiation in buccal mucosal fibroblasts. J. Formos. Med. Assoc. 2018, 117, 791–797. [Google Scholar] [CrossRef]
- Byun, Y.; Choi, Y.C.; Jeong, Y.; Lee, G.; Yoon, S.; Jeong, Y.; Yoon, J.; Baek, K. MiR-200c downregulates HIF-1alpha and inhibits migration of lung cancer cells. Cell. Mol. Biol. Lett. 2019, 24, 28. [Google Scholar] [CrossRef] [Green Version]
- Li, L.; Li, B.; Chen, D.; Liu, L.; Huang, C.; Lu, Z.; Lun, L.; Wan, X. miR-139 and miR-200c regulate pancreatic cancer endothelial cell migration and angiogenesis. Oncol. Rep. 2015, 34, 51–58. [Google Scholar] [CrossRef] [Green Version]
- Maolakuerban, N.; Azhati, B.; Tusong, H.; Abula, A.; Yasheng, A.; Xireyazidan, A. MiR-200c-3p inhibits cell migration and invasion of clear cell renal cell carcinoma via regulating SLC6A1. Cancer Biol. Ther. 2018, 19, 282–291. [Google Scholar] [CrossRef] [Green Version]
- Shao, X.L.; Chen, Y.; Gao, L. MiR-200c suppresses the migration of retinoblastoma cells by reversing epithelial mesenchymal transition. Int. J. Ophthalmol. 2017, 10, 1195–1202. [Google Scholar]
- Liu, L.; Qiu, M.; Tan, G.; Liang, Z.; Qin, Y.; Chen, L.; Chen, H.; Liu, J. miR-200c inhibits invasion, migration and proliferation of bladder cancer cells through down-regulation of BMI-1 and E2F3. J. Transl. Med. 2014, 12, 305. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Guo, K.; Wang, J.; Shu, L.; Zhou, G. MiR-200c promotes papillary thyroid cancer cell proliferation, migration, and invasion by downregulating PTEN. Tissue Cell 2021, 73, 101647. [Google Scholar] [CrossRef] [PubMed]
- Lu, Z.; Du, L.; Liu, R.; Di, R.; Zhang, L.; Ma, Y.; Li, Q.; Liu, E.; Chu, M.; Wei, C. MiR-378 and BMP-Smad can influence the proliferation of sheep myoblast. Gene 2018, 674, 143–150. [Google Scholar] [CrossRef] [PubMed]
- Zeng, M.; Zhu, L.; Li, L.; Kang, C. miR-378 suppresses the proliferation, migration and invasion of colon cancer cells by inhibiting SDAD1. Cell. Mol. Biol. Lett. 2017, 22, 12. [Google Scholar] [CrossRef] [Green Version]
- Bartel, D.P. MicroRNAs: Target recognition and regulatory functions. Cell 2009, 136, 215–233. [Google Scholar] [CrossRef] [Green Version]
- Fabian, M.R.; Sonenberg, N.; Filipowicz, W. Regulation of mRNA translation and stability by microRNAs. Annu. Rev. Biochem. 2010, 79, 351–379. [Google Scholar] [CrossRef] [Green Version]
- Tang, B.L. ADAMTS: A novel family of extracellular matrix proteases. Int. J. Biochem. Cell Biol. 2001, 33, 33–44. [Google Scholar] [CrossRef]
- Haraguchi, N.; Ohara, N.; Koseki, J.; Takahashi, H.; Nishimura, J.; Hata, T.; Mizushima, T.; Yamamoto, H.; Ishii, H.; Doki, Y.; et al. High expression of ADAMTS5 is a potent marker for lymphatic invasion and lymph node metastasis in colorectal cancer. Mol. Clin. Oncol. 2017, 6, 130–134. [Google Scholar] [CrossRef] [Green Version]
- Gu, J.; Chen, J.; Feng, J.; Liu, Y.; Xue, Q.; Mao, G.; Gai, L.; Lu, X.; Zhang, R.; Cheng, J.; et al. Overexpression of ADAMTS5 can regulate the migration and invasion of non-small cell lung cancer. Tumour Biol. 2016, 37, 8681–8689. [Google Scholar] [CrossRef]
- Held-Feindt, J.; Paredes, E.B.; Blomer, U.; Seidenbecher, C.; Stark, A.M.; Mehdorn, H.M.; Mentlein, R. Matrix-degrading proteases ADAMTS4 and ADAMTS5 (disintegrins and metalloproteinases with thrombospondin motifs 4 and 5) are expressed in human glioblastomas. Int. J. Cancer 2006, 118, 55–61. [Google Scholar] [CrossRef]
- Akcora-Yildiz, D.; Yukselten, Y.; Sunguroglu, M.; Ugur, H.C.; Sunguroglu, A. IL-33 induces ADAMTS5 expression and cell migration in glioblastoma multiforme. Med. Oncol. 2022, 39, 22. [Google Scholar] [CrossRef]
- Huang, J.; Sun, Y.; Chen, H.; Liao, Y.; Li, S.; Chen, C.; Yang, Z. ADAMTS5 acts as a tumor suppressor by inhibiting migration, invasion and angiogenesis in human gastric cancer. Gastric Cancer 2019, 22, 287–301. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Li, C.; Xiong, Y.; Yang, X.; Wang, L.; Zhang, S.; Dai, N.; Li, M.; Ren, T.; Yang, Y.; Zhou, S.F.; et al. Lost expression of ADAMTS5 protein associates with progression and poor prognosis of hepatocellular carcinoma. Drug Des. Dev. Ther. 2015, 9, 1773–1783. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Endo, T. Molecular mechanisms of skeletal muscle development, regeneration, and osteogenic conversion. Bone 2015, 80, 2–13. [Google Scholar] [CrossRef] [PubMed]
- Zhang, Z.K.; Li, J.; Guan, D.; Liang, C.; Zhuo, Z.; Liu, J.; Lu, A.; Zhang, G.; Zhang, B.T. A newly identified lncRNA MAR1 acts as a miR-487b sponge to promote skeletal muscle differentiation and regeneration. J. Cachexia Sarcopenia Muscle 2018, 9, 613–626. [Google Scholar] [CrossRef] [Green Version]
- Yuan, Y.; Shi, X.E.; Liu, Y.G.; Yang, G.S. FoxO1 regulates muscle fiber-type specification and inhibits calcineurin signaling during C2C12 myoblast differentiation. Mol. Cell. Biochem. 2011, 348, 77–87. [Google Scholar] [CrossRef]
- Wang, H.; Ma, M.; Li, Y.; Liu, J.; Sun, C.; Liu, S.; Ma, Y.; Yan, Y.; Tang, Z.; Shen, S.; et al. miR-183 and miR-96 orchestrate both glucose and fat utilization in skeletal muscle. EMBO Rep. 2021, 22, e52247. [Google Scholar] [CrossRef]
- Zhang, D.; Li, Y.; Yao, X.; Wang, H.; Zhao, L.; Jiang, H.; Yao, X.; Zhang, S.; Ye, C.; Liu, W.; et al. miR-182 Regulates Metabolic Homeostasis by Modulating Glucose Utilization in Muscle. Cell Rep. 2016, 16, 757–768. [Google Scholar] [CrossRef] [Green Version]
- Li, D.; Wang, Y.; Jin, X.; Hu, D.; Xia, C.; Xu, H.; Hu, J. NK cell-derived exosomes carry miR-207 and alleviate depression-like symptoms in mice. J. Neuroinflammat. 2020, 17, 126. [Google Scholar] [CrossRef] [PubMed] [Green Version]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Liu, Y.; Yao, Y.; Zhang, Y.; Yan, C.; Yang, M.; Wang, Z.; Li, W.; Li, F.; Wang, W.; Yang, Y.; et al. MicroRNA-200c-5p Regulates Migration and Differentiation of Myoblasts via Targeting Adamts5 in Skeletal Muscle Regeneration and Myogenesis. Int. J. Mol. Sci. 2023, 24, 4995. https://doi.org/10.3390/ijms24054995
Liu Y, Yao Y, Zhang Y, Yan C, Yang M, Wang Z, Li W, Li F, Wang W, Yang Y, et al. MicroRNA-200c-5p Regulates Migration and Differentiation of Myoblasts via Targeting Adamts5 in Skeletal Muscle Regeneration and Myogenesis. International Journal of Molecular Sciences. 2023; 24(5):4995. https://doi.org/10.3390/ijms24054995
Chicago/Turabian StyleLiu, Yanwen, Yilong Yao, Yongsheng Zhang, Chao Yan, Mingsha Yang, Zishuai Wang, Wangzhang Li, Fanqinyu Li, Wei Wang, Yalan Yang, and et al. 2023. "MicroRNA-200c-5p Regulates Migration and Differentiation of Myoblasts via Targeting Adamts5 in Skeletal Muscle Regeneration and Myogenesis" International Journal of Molecular Sciences 24, no. 5: 4995. https://doi.org/10.3390/ijms24054995