Skip to main content
Log in

Downregulation of miR-27a* and miR-532-5p and Upregulation of miR-146a and miR-155 in LPS-induced RAW264.7 Macrophage Cells

  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

MicroRNAs (miRNAs) are short non-coding RNAs that are involved in the epigenetic regulation of cellular processes. To identify more miRNAs which are involved in the macrophage inflammatory response to lipopolysaccharide (LPS) stimulation and dissect the mechanisms more clearly, microRNA profiling of LPS-treated RAW264.7 macrophage cells was performed by initial high-throughput array-based screen and further real-time RT-PCR validation; bioinformatics approaches were used to analyze the target genes of the differentially expressed miRNAs. Compared to the untreated control, two microRNAs (miR-146a and miR-155) with more than twofold higher expression and two microRNAs (miR-27a* and miR-532-5p) with twofold lower expression were detected by array-based screen, which can be validated by qRT-PCR, and more than 1,000 candidate target genes were detected by at least of one of four different algorithms (TargetScan, PicTar, miRDB, and microRNA.org); with gene ontology classification, we were able to correlate the upregulation and downregulation of miRNA to the differential expression of inflammation-related candidate target gene during LPS-induced inflammation. Our findings may provide the basic information for the precise roles of miRNAs in the macrophage inflammatory response to LPS stimulation in the future.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

REFERENCES

  1. Abbott, A.L., E. Alvarez-Saavedra, E.A. Miska, N.C. Lau, D.P. Bartel, H.R. Horvitz, and V. Ambros. 2005. The let-7 microRNA family members mir-48, mir-84, and mir-241 function together to regulate developmental timing in Caenorhabditis elegans. Developmental Cell 9: 403–414.

    Article  PubMed  CAS  Google Scholar 

  2. Androulidaki, A., D. Iliopoulos, A. Arranz, C. Doxaki, S. Schworer, V. Zacharioudaki, A.N. Margioris, P.N. Tsichlis, and C. Tsatsanis. 2009. The kinase Akt1 controls macrophage response to lipopolysaccharide by regulating microRNA. Immunity 31: 220–231.

    Article  PubMed  CAS  Google Scholar 

  3. Chen, C.Z., L. Li, H.F. Lodish, and D.P. Bartel. 2004. MicroRNAs modulate hematopoietic lineage differentiation. Science 303: 83–86.

    Article  PubMed  CAS  Google Scholar 

  4. Cheng, A.M., M.W. Byrom, J. Shelton, and L.P. Ford. 2005. Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis. Nucleic Acids Research 33: 1290–1297.

    Article  PubMed  CAS  Google Scholar 

  5. Dong, W., Y. Liu, J. Peng, L. Chen, T. Zou, H. Xiao, Z. Liu, W. Li, Y. Bu, and Y. Qi. 2006. The IRAK-BCL10-MALT1-TRAF6-TAK1 cascade mediates signaling to NF-κB from toll-like receptor 4. The Journal of Biological Chemistry 281: 26029–26040.

    Article  PubMed  CAS  Google Scholar 

  6. Janeway, C.A., and R. Medzhitov. 2002. Innate immune recognition. Annual Review of Immunology 20: 197–216.

    Article  PubMed  CAS  Google Scholar 

  7. Lei, M., H. Jiao, T. Liu, L. Du, Y. Cheng, D. Zhang, Y. Hao, C. Man, and F. Wang. 2011. siRNA targeting mCD14 inhibits TNF-α, MIP-2, and IL-6 secretion and NO production from LPS-induced RAW264.7 cells. Applied Microbiology and Biotechnology 92: 115–124.

    Article  PubMed  CAS  Google Scholar 

  8. Lewis, B.P., C.B. Burge, and D.P. Bartel. 2005. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120: 15–20.

    Article  PubMed  CAS  Google Scholar 

  9. Liu, J., S. Batkai, P. Pacher, J. Harvey-White, J.A. Wagner, B.F. Cravatt, B. Gao, and G. Kunos. 2003. Lipopolysaccharide induces anandamide synthesis in macrophages via CD14/MAPK/Phosphoinositide3-kinase/NF-κB independently of platelet-activating factor. The Journal of Biological Chemistry 278: 45034–45039.

    Article  PubMed  CAS  Google Scholar 

  10. Lu, Y., J.M. Thomson, H.Y. Wong, S.M. Hammond, and B.L. Hogan. 2007. Transgenic overexpression of the microRNA miR-17-92 cluster promotes proliferation and inhibits differentiation of lung epithelial progenitor cells. Developmental Biology 310: 442–453.

    Article  PubMed  CAS  Google Scholar 

  11. Lu, L.F., T.H. Thai, D.P. Calado, A. Chaudhry, M. Kubo, K. Tanaka, G.B. Loeb, H. Lee, A. Yoshimura, K. Rajewsky, and A.Y. Rudensky. 2009. Foxp3-dependent microRNA155 confers competitive fitness to regulatory T cells by targeting SOCS1 protein. Immunity 30: 80–91.

    Article  PubMed  CAS  Google Scholar 

  12. Mallory, A.C., and H. Vaucheret. 2004. MicroRNAs: something important between the genes. Current Opinion in Plant Biology 7: 120–125.

    Article  PubMed  CAS  Google Scholar 

  13. Maris, N.A., M.C. Dessing, A.F. de Vos, P. Bresser, J.S. van der Zee, H.M. Jansen, C.A. Spek, and T. van der Poll. 2006. Toll-like receptor mRNA levels in alveolar macrophages after inhalation of endotoxin. European Respiratory Journal 28: 622–626.

    Article  PubMed  CAS  Google Scholar 

  14. Molnár, A., F. Schwach, D.J. Studholme, E.C. Thuenemann, and D.C. Baulcombe. 2007. miRNAs control gene expression in the single-cell alga Chlamydomonas reinhardtii. Nature 447: 1126–1129.

    Article  PubMed  Google Scholar 

  15. Matsuguchi, T., A. Masuda, K. Sugimoto, Y. Nagai, and Y. Yoshikai. 2003. JNK-interacting protein 3 associates with toll-like receptor 4 and is involved in LPS-mediated JNK activation. The EMBO Journal 22: 4455–4464.

    Article  PubMed  CAS  Google Scholar 

  16. O’Connell, R.M., A.A. Chaudhuri, D.S. Rao, and D. Baltimore. 2009. Inositol phosphatase SHIP1 is a primary target of miR-155. Proceedings of the National Academy of Sciences of the United States of America 106: 7113–7118.

    Article  PubMed  Google Scholar 

  17. Sarkar, S., J. Han, K.S. Sinsimer, B. Liao, R.L. Foster, G. Brewer, and S. Pestka. 2011. RNA-binding protein AUF1 regulates lipopolysaccharide-induced IL10 expression by activating IκB kinase complex in monocytes. Molecular and Cellular Biology 31: 602–615.

    Article  PubMed  CAS  Google Scholar 

  18. Schmittgen, T.D., and K.J. Livak. 2008. Analyzing real-time PCR data by the comparative C(T) method. Nature Protocols 3: 1101–1108.

    Article  PubMed  CAS  Google Scholar 

  19. Taganov, K.D., M.P. Boldin, K.J. Chang, and D. Baltimore. 2006. NF-κB dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proceedings of the National Academy of Sciences of the United States of America 103: 12481–12486.

    Article  PubMed  CAS  Google Scholar 

  20. Tili, E., J.J. Michaille, A. Cimino, S. Costinean, C.D. Dumitru, B. Adair, M. Fabbri, H. Alder, C.G. Liu, G.A. Calin, and C.M. Croce. 2007. Modulation of miR-155 and miR-125b levels following lipopolysaccharide/TNF-α stimulation and their possible roles in regulation the response to entodoxin shock. Journal of Immunology 179: 5082–5089.

    CAS  Google Scholar 

  21. Zhan, M., C.P. Miller, T. Papayannopoulou, G. Stamatoyannopoulos, and C.Z. Song. 2007. MicroRNA expression dynamics during murine and human erythroid differentiation. Experimental Hematology 35: 1015–1025.

    Article  PubMed  CAS  Google Scholar 

  22. Zhong, H., H.R. Wang, S. Yang, J.H. Zhong, T. Wang, C. Wang, and F.Y. Chen. 2010. Targeting Smad4 links microRNA-146a to the TGF-β pathway during retinoid acid induction in acute promyelocytic leukemia cell line. International Journal of Hematology 92: 129–135.

    Article  PubMed  CAS  Google Scholar 

Download references

ACKNOWLEDGMENTS

This study was financially supported by the Major National Science and Technology Specific Projects (no. 2009ZX08007-009B) and the National High Technology Research and Development Program of China (2011AA100302).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Fengyang Wang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cheng, Y., Kuang, W., Hao, Y. et al. Downregulation of miR-27a* and miR-532-5p and Upregulation of miR-146a and miR-155 in LPS-induced RAW264.7 Macrophage Cells. Inflammation 35, 1308–1313 (2012). https://doi.org/10.1007/s10753-012-9443-8

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10753-012-9443-8

KEY WORDS

Navigation