Spinal circRNA-9119 Suppresses Nociception by Mediating the miR-26a-TLR3 Axis in a Bone Cancer Pain Mouse Model

  • Zhongqi Zhang
  • Xiaoxia Zhang
  • Yanjing Zhang
  • Jiyuan Li
  • Zumin XingEmail author
  • Yiwen ZhangEmail author


Altered expression of circular RNA (circRNA) is recognized as a contributor to malignant pain where microRNA (miRNA) exerts an essential effect. We generated a murine model for bone malignancy pain in which 2472 osteolytic sarcoma cells were injected into the femurs of mice. CircRNA microarray and quantitative PCR (qPCR) and revealed that circ9119 expression was repressed in the spinal cord of bone malignancy pain model mice, which is the first relay site involved in the transmission of nociceptive information to the cerebrum of mice that receive spinal analgesics for malignancy pain. Overexpression of circ9119 by plasmid injection in the model mice reduced progressive thermal hyperalgesia and mechanical hyperalgesia. Bioinformatics prediction and dual-luciferase reporter assay showed that circ9119 functions as a sponge of miR-26a, which targets the TLR3 3′-untranslated region. Furthermore, expression of miR-26a was elevated and TLR3 level was repressed in bone malignancy pain model mice, which were counteracted by circ9119 in the spinal cord of tumor-bearing mice. Moreover, excessive expression of miR-26a was involved in the recovery of mice from progressive thermal hyperalgesia and mechanical hyperalgesia triggered via circ9119. TLR3 knockdown in bone malignancy pain model mice thoroughly impaired pain in the initial stages and reduced the effects of circ9119 on hyperalgesia. Our research findings indicate that targeting the circ9119-miR-26a-TLR3 axis may be a promising analgesic strategy to manage malignancy pain.


Bone cancer pain Hyperalgesia Circular RNA-9119 miR-26a TLR3 



This work was supported by the Key Specialist Project of Clinical Medicine of Foshan City (Grant Number FSZDZK135049); Distinguished Youth Talent Fund Project of the First Medical Science Center of Foshan City in 2018; and Guangdong Medical Research Fund Project in 2019 (Grant Number A2019045). We thank Edanz Group ( for editing a draft of this manuscript.

Compliance with Ethical Standards

This study was approved by the Committee on the Ethics of Animal Experiments of Shunde Hospital of Southern Medical University.

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. Chen I, Chen CY, Chuang TJ (2015) Biogenesis, identification, and function of exonic circular RNAs. Wiley Interdiscip Rev RNA 6:563–579. CrossRefPubMedPubMedCentralGoogle Scholar
  2. Chen W, Lu Z (2017) Upregulated TLR3 promotes neuropathic pain by regulating autophagy in rat with L5 spinal nerve ligation model. Neurochem Res 42:634–643. CrossRefPubMedGoogle Scholar
  3. Coleman RE (1997) Skeletal complications of malignancy cancer: interdisciplinary. Int J Am Cancer Soc 80:1588–1594Google Scholar
  4. Coyle N, Adelhardt J, Foley KM, Portenoy RK (1990) Character of terminal illness in the advanced cancer patient: pain and other symptoms during the last four weeks of life. J Pain Symptom Manag 5:83–93CrossRefGoogle Scholar
  5. Ding K et al (2017) MiR-26a performs converse roles in proliferation and metastasis of different gastric cancer cells via regulating of PTEN expression. Pathol Res Pract 213:467–475. CrossRefPubMedGoogle Scholar
  6. Ebbesen KK, Kjems J, Hansen TB (2016) Circular RNAs: identification, biogenesis and function. Biochim Biophys Acta 1859:163–168. CrossRefPubMedGoogle Scholar
  7. Elramah S et al (2017) Spinal miRNA-124 regulates synaptopodin and nociception in an animal model of bone cancer pain. Sci Rep 7:10949CrossRefGoogle Scholar
  8. Fujii T, Shimada K, Nakai T, Ohbayashi C (2018) MicroRNAs in smoking-related carcinogenesis: biomarkers, functions, and therapy. J Clin Med:7. CrossRefGoogle Scholar
  9. Gandla J, Lomada SK, Lu J, Kuner R, Bali KK (2017) miR-34c-5p functions as pronociceptive microRNA in cancer pain by targeting Cav2. 3 containing calcium channels. Pain 158:1765CrossRefGoogle Scholar
  10. Hansen TB, Jensen TI, Clausen BH, Bramsen JB, Finsen B, Damgaard CK, Kjems J (2013) Natural RNA circles function as efficient microRNA sponges. Nature 495:384CrossRefGoogle Scholar
  11. Hargreaves K, Dubner R, Brown F, Flores C, Joris J (1988) A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia. Pain 32:77–88CrossRefGoogle Scholar
  12. Hayes J, Peruzzi PP, Lawler S (2014) MicroRNAs in cancer: biomarkers, functions and therapy. Trends Mol Med 20:460–469. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Hsieh CH et al (2017) Knockout of toll-like receptor impairs nerve regeneration after a crush injury. Oncotarget 8:80741–80756. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Jeck WR et al. (2013) Circular RNAs are abundant, conserved, and associated with ALU repeats RNA 19:141-157 doi: CrossRefGoogle Scholar
  15. Jiang C et al (2014b) MicroRNA-26a negatively regulates toll-like receptor 3 expression of rat macrophages and ameliorates pristane induced arthritis in rats. Arthritis Research & Therapy 16:R9CrossRefGoogle Scholar
  16. Jiang D-S, Wang Y-W, Jiang J, Li S-M, Liang S-Z, Fang H-Y (2014a) MicroRNA-26a involved in toll-like receptor 9-mediated lung cancer growth and migration. Int J Mol Med 34:307–312CrossRefGoogle Scholar
  17. Lacagnina MJ, Watkins LR, Grace PM (2018) Toll-like receptors and their role in persistent pain. Pharmacol Ther 184:145–158. CrossRefPubMedGoogle Scholar
  18. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method methods 25:402–408CrossRefGoogle Scholar
  19. Lopez-Urrutia E et al (2017) MiR-26a downregulates retinoblastoma in colorectal cancer. Tumour Biol 39:1010428317695945. CrossRefPubMedGoogle Scholar
  20. Luger NM et al (2001) Osteoprotegerin diminishes advanced bone cancer pain. Cancer Res 61:4038–4047PubMedGoogle Scholar
  21. Mei X-P, Zhou Y, Wang W, Tang J, Wang W, Zhang H, Xu LX, Li YQ (2011) Ketamine depresses toll-like receptor 3 signaling in spinal microglia in a rat model of neuropathic pain. Neurosignals 19:44–53CrossRefGoogle Scholar
  22. Mercadante S (1997) Malignant bone pain: pathophysiology and treatment. Pain 69:1–18CrossRefGoogle Scholar
  23. Njoo C, Heinl C, Kuner R (2014) In vivo SiRNA transfection and gene knockdown in spinal cord via rapid noninvasive lumbar intrathecal injections in mice JoVE (Journal of Visualized Experiments):e51229Google Scholar
  24. Portenoy RK, Lesage P (1999) Management of cancer pain. Lancet 353:1695–1700CrossRefGoogle Scholar
  25. Qi J et al (2011) Painful pathways induced by TLR stimulation of dorsal root ganglion neurons. J Immunol (Baltimore, Md : 1950) 186:6417–6426. CrossRefGoogle Scholar
  26. Reuland SN et al (2013) MicroRNA-26a is strongly downregulated in melanoma and induces cell death through repression of silencer of death domains (SODD). J Investig Dermatol 133:1286–1293CrossRefGoogle Scholar
  27. Rizzo M et al (2017) Discovering the miR-26a-5p targetome in prostate cancer cells. J Cancer 8:2729–2739. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Rybak-Wolf A, Stottmeister C, Glažar P, Jens M, Pino N, Giusti S, Hanan M, Behm M, Bartok O, Ashwal-Fluss R, Herzog M, Schreyer L, Papavasileiou P, Ivanov A, Öhman M, Refojo D, Kadener S, Rajewsky N (2015) Circular RNAs in the mammalian brain are highly abundant, conserved, and dynamically expressed. Mol Cell 58:870–885. CrossRefPubMedGoogle Scholar
  29. Sabino MAC et al. (2002) Simultaneous reduction in cancer pain, bone destruction, and tumor growth by selective inhibition of cyclooxygenase-2 62:7343-7349Google Scholar
  30. Schwei MJ, Honore P, Rogers SD, Salak-Johnson JL, Finke MP, Ramnaraine ML, Clohisy DR, Mantyh PW (1999) Neurochemical and cellular reorganization of the spinal cord in a murine model of bone cancer pain. J Neurosci 19:10886–10897CrossRefGoogle Scholar
  31. Tétreault P, Dansereau M-A, Doré-Savard L, Beaudet N, Sarret P (2011) Weight bearing evaluation in inflammatory, neuropathic and cancer chronic pain in freely moving rats. Physiol Behav 104:495–502CrossRefGoogle Scholar
  32. Zhang L et al (2018a) CircRNA-9119 regulates the expression of prostaglandin-endoperoxide synthase 2 (PTGS2) by sponging miR-26a in the endometrial epithelial cells of dairy goat. Reprod Fertil Dev. CrossRefGoogle Scholar
  33. Zhang L et al (2018b) CircRNA-9119 regulates the expression of prostaglandin-endoperoxide synthase 2 (PTGS2) by sponging miR-26a in the endometrial epithelial cells of dairy goat. 30:1759–1769Google Scholar
  34. Zhang Y et al (2015) MicroRNA-26a prevents endothelial cell apoptosis by directly targeting TRPC6 in the setting of atherosclerosis. Sci Rep 5:9401CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019
corrected publication 2019

Authors and Affiliations

  1. 1.Department of AnesthesiologyShunde Hospital of Southern Medical UniversityFoshanChina

Personalised recommendations