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MicroRNA-let-7a promotes E2F-mediated cell proliferation and NFκB activation in vitro

Cellular & Molecular Immunology volume 11, pages 79–83 (2014)Cite this article

Abstract

Epigenetic factors, including altered microRNA (miRNA) expression, may contribute to aberrant immune cell function in systemic lupus erythematosus (SLE). MiRNA-let-7a (let-7a) has been shown to directly alter cell cycle progression and proinflammatory cytokine production. Due to the crucial role of let-7a in cell division and inflammation, we investigated let-7a-mediated proliferation and NFκB translocation in J774A.1 macrophages and MES 13 mesangial cells in vitro. In immune-stimulated cells transfected with let-7a, cell proliferation was significantly increased over time. There was a significant increase in the number of immune-stimulated cells in S and G2 phases. Immune-stimulated cells overexpressing let-7a had increased nuclear translocation of NFκB. Bioinformatical analysis revealed that the E2F family, critical regulators of the G1–S transition, has potential binding sites for let-7a in their mRNA transcripts. Let-7a overexpression significantly increased the expression of the cell cycle activator E2F2 and increased retinoblastoma protein (Rb) phosphorylation in immune-stimulated cells. The cell cycle inhibitor E2F5 was significantly decreased in let-7a-transfected cells that were immune-stimulated. Bioinformatical analysis revealed E2F2 and NFκB are transcription factors predicted to regulate the let-7a promoter. We analyzed transcriptional regulation of let-7a by real-time RT-PCR using chromatin immunoprecipitation with E2F2 and NFκB antibodies. There was an increase in E2F2 and NFκB binding in DNA enriched for the let-7a promoter in immune-stimulated cells. Silencing E2F2 or NFκB significantly decreased let-7a expression and IL-6 production in immune-stimulated cells. Taken together, our results suggest that overexpression of let-7a may contribute to hyperplasia and the proinflammatory response in SLE.

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Acknowledgements

We wish to thank Melissa Makris for her assistance with flow data analyses. This work was supported by a grant from the National Institutes of Health/National Institute of Allergy and Infectious Diseases (R15 AR062883).

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Authors and Affiliations

  1. Department of Biomedical Sciences & Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA

    Cristen B Chafin, Nicole L Regna & Christopher M Reilly

  2. Department of Pathology/Comparative Medicine, Wake Forest University Primate Center, Wake Forest School of Medicine, Winston-Salem, NC, USA

    David L Caudell

  3. Edward Via College of Osteopathic Medicine, Blacksburg, VA, USA

    Christopher M Reilly

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Correspondence to Cristen B Chafin.

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Chafin, C., Regna, N., Caudell, D. et al. MicroRNA-let-7a promotes E2F-mediated cell proliferation and NFκB activation in vitro. Cell Mol Immunol 11, 79–83 (2014). https://doi.org/10.1038/cmi.2013.51

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