Neuroscience Bulletin

, Volume 28, Issue 2, pp 165–172 | Cite as

Formaldehyde up-regulates TRPV1 through MAPK and PI3K signaling pathways in a rat model of bone cancer pain

  • Ying Han
  • Yan Li
  • Xing Xiao
  • Jia Liu
  • Xiang-Ling Meng
  • Feng-Yu Liu
  • Guo-Gang Xing
  • You Wan
Original Article



Our previous study showed that tumor tissue-derived formaldehyde at low concentrations plays an important role in bone cancer pain through activating transient receptor potential vanilloid subfamily member 1 (TRPV1). The present study further explored whether this tumor tissue-derived endogenous formaldehyde regulates TRPV1 expression in a rat model of bone cancer pain, and if so, what the possible signal pathways are during the development of this type of pain.


A rat model of bone cancer pain was established by injecting living MRMT-1 tumor cells into the tibia. The formaldehyde levels were determined by high performance liquid chromatography, and the expression of TRPV1 was examined with Western blot and RT-PCR. In primary cultured dorsal root ganglion (DRG) neurons, the expression of TRPV1 was assessed after treatment with 100 μmol/L formaldehyde with or without pre-addition of PD98059 [an inhibitor for extracellular signal-regulated kinase], SB203580 (a p38 inhibitor), SP600125 [an inhibitor for c-Jun N-terminal kinase], BIM [a protein kinase C (PKC) inhibitor] or LY294002 [a phosphatidylinositol 3-kinase (PI3K) inhibitor].


In the rat model of bone cancer pain, formaldehyde concentration increased in blood plasma, bone marrow and the spinal cord. TRPV1 protein expression was also increased in the DRG. In primary cultured DRG neurons, 100 μmol/L formaldehyde significantly increased the TRPV1 expression level. Pre-incubation with PD98059, SB203580, SP600125 or LY294002, but not BIM, inhibited the formaldehyde-induced increase of TRPV1 expression.


Formaldehyde at a very low concentration up-regulates TRPV1 expression through mitogen-activated protein kinase and PI3K, but not PKC, signaling pathways. These results further support our previous finding that TRPV1 in peripheral afferents plays a role in bone cancer pain.


formaldehyde TRPV1 cancer pain mitogen-activated protein kinase phosphatidylinositol 3-kinase 


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  1. [1]
    Portenoy RK, Payne D, Jacobsen P. Breakthrough pain: characteristics and impact in patients with cancer pain. Pain 1999, 81: 129–134.PubMedCrossRefGoogle Scholar
  2. [2]
    Ghilardi JR, Röhrich H, Lindsay TH, Sevcik MA, Schwei MJ, Kubota K, et al. Selective blockade of the capsaicin receptor TRPV1 attenuates bone cancer pain. J Neurosci 2005, 25: 3126–3131.PubMedCrossRefGoogle Scholar
  3. [3]
    Kalasz H. Biological role of formaldehyde, and cycles related to methylation, demethylation, and formaldehyde production. Mini Rev Med Chem 2003, 3: 175–192.PubMedCrossRefGoogle Scholar
  4. [4]
    Ebeler SE, Clifford AJ, Shibamoto T. Quantitative analysis by gas chromatography of volatile carbonyl compounds in expired air from mice and human. J Chromatogr B Biomed Sci Appl 1997, 702: 211–215.PubMedCrossRefGoogle Scholar
  5. [5]
    Spanel P, Smith D, Holland TA, Al Singary W, Elder JB. Analysis of formaldehyde in the headspace of urine from bladder and prostate cancer patients using selected ion flow tube mass spectrometry. Rapid Commun Mass Spectrom 1999, 13: 1354–1359.PubMedCrossRefGoogle Scholar
  6. [6]
    Sabino MA, Mantyh PW. Pathophysiology of bone cancer pain. J Support Oncol 2005, 3: 15–24.PubMedGoogle Scholar
  7. [7]
    Thorndike J, Beck WS. Production of formaldehyde from N5-methyltetrahydrofolate by normal and leukemic leukocytes. Cancer Res 1977, 37: 1125–1132.PubMedGoogle Scholar
  8. [8]
    Tong Z, Luo W, Wang Y, Yang F, Han Y, Li H, et al. Tumor tissuederived formaldehyde and acidic microenvironment synergistically induce bone cancer pain. PLoS One 2010, 5: e10234.PubMedCrossRefGoogle Scholar
  9. [9]
    Pei L, Lin CY, Dai JP, Yin GF. Facial pain induces the alteration of transient receptor potential vanilloid receptor 1 expression in rat trigeminal ganglion. Neurosci Bull 2007, 23: 92–100.PubMedCrossRefGoogle Scholar
  10. [10]
    Luo H, Cheng J, Han JS, Wan Y. Change of vanilloid receptor 1 expression in dorsal root ganglion and spinal dorsal horn during inflammatory nociception induced by complete Freund’s adjuvant in rats. Neuroreport 2004, 15: 655–658.PubMedCrossRefGoogle Scholar
  11. [11]
    Christoph T, Grunweller A, Mika J, Schäfer MK, Wade EJ, Weihe E, et al. Silencing of vanilloid receptor TRPV1 by RNAi reduces neuropathic and visceral pain in vivo. Biochem Biophys Res Commun 2006, 350: 238–243.PubMedCrossRefGoogle Scholar
  12. [12]
    Niiyama Y, Kawamata T, Yamamoto J, Omote K, Namiki A. Bone cancer increases transient receptor potential vanilloid subfamily 1 expression within distinct subpopulations of dorsal root ganglion neurons. Neuroscience 2007, 148: 560–572.PubMedCrossRefGoogle Scholar
  13. [13]
    Yu L, Yang F, Luo H, Liu FY, Han JS, Xing GG, et al. The role of TRPV1 in different subtypes of dorsal root ganglion neurons in rat chronic inflammatory nociception induced by complete Freund’s adjuvant. Mol Pain 2008, 4: 61.PubMedCrossRefGoogle Scholar
  14. [14]
    Niiyama Y, Kawamata T, Yamamoto J, Furuse S, Namiki A. SB366791, a TRPV1 antagonist, potentiates analgesic effects of systemic morphine in a murine model of bone cancer pain. Br J Anaesth 2009, 102: 251–258.PubMedCrossRefGoogle Scholar
  15. [15]
    Du Y, Xiao Y, Lu ZM, Ding J, Xie F, Fu H, et al. Melittin activates TRPV1 receptors in primary nociceptive sensory neurons via the phospholipase A2 cascade pathways. Biochem Biophys Res Commun 2011, 408: 32–37.PubMedCrossRefGoogle Scholar
  16. [16]
    Xia R, Samad TA, Btesh J, Jiang LH, Kays I, Stjernborg L, et al. TRPV1 signaling: mechanistic understanding and therapeutic potential. Curr Top Med Chem 2011, 11: 2180–2191.PubMedCrossRefGoogle Scholar
  17. [17]
    Tian LJ, Du Y, Xiao Y, Lv ZM, Yu YQ, Cui XY, et al. Mediating roles of the vanilloid receptor TRPV1 in activation of rat primary afferent nociceptive neurons by formaldehyde. Acta Physiol Sin 2009, 61: 404–416.Google Scholar
  18. [18]
    Svensson C, Part K, Kunnis-Beres K, Kaldmäe M, Fernaeus SZ, Land T. Pro-survival effects of JNK and p38 MAPK pathways in LPS-induced activation of BV-2 cells. Biochem Biophys Res Commun 2011, 406: 488–492.PubMedCrossRefGoogle Scholar
  19. [19]
    Doya H, Ohtori S, Fujitani M, Saito T, Hata K, Ino H, et al. c-Jun N-terminal kinase activation in dorsal root ganglion contributes to pain hypersensitivity. Biochem Biophys Res Commun 2005, 335: 132–138.PubMedCrossRefGoogle Scholar
  20. [20]
    Ji RR, Kawasaki Y, Zhuang ZY, Wen YR, Zhang YQ. Protein kinases as potential targets for the treatment of pathological pain. Handb Exp Pharmacol 2007: 359–389.Google Scholar
  21. [21]
    Ji RR, Strichartz G. Cell signaling and the genesis of neuropathic pain. Sci STKE 2004, 2004: E14.CrossRefGoogle Scholar
  22. [22]
    Ji RR, Samad TA, Jin SX, Schmoll R, Woolf CJ. p38 MAPK activation by NGF in primary sensory neurons after inflammation increases TRPV1 levels and maintains heat hyperalgesia. Neuron 2002, 36: 57–68.PubMedCrossRefGoogle Scholar
  23. [23]
    Morenilla-Palao C, Planells-Cases R, Garcia-Sanz N, Ferrer-Montiel A. Regulated exocytosis contributes to protein kinase C potentiation of vanilloid receptor activity. J Biol Chem 2004, 279: 25665–25672.PubMedCrossRefGoogle Scholar
  24. [24]
    Zhuang ZY, Xu H, Clapham DE, Ji RR. Phosphatidylinositol 3-kinase activates ERK in primary sensory neurons and mediates inflammatory heat hyperalgesia through TRPV1 sensitization. J Neurosci 2004, 24: 8300–8309.PubMedCrossRefGoogle Scholar
  25. [25]
    Amadesi S, Cottrell GS, Divino L, Chapman K, Grady EF, Bautista F, et al. Protease-activated receptor 2 sensitizes TRPV1 by protein kinase C epsilon- and A-dependent mechanisms in rats and mice. J Physiol 2006, 575(Pt 2): 555–571.PubMedCrossRefGoogle Scholar
  26. [26]
    Zhang H, Cang CL, Kawasaki Y, Liang LL, Zhang YQ, Ji RR, et al. Neurokinin-1 receptor enhances TRPV1 activity in primary sensory neurons via PKC epsilon: a novel pathway for heat hyperalgesia. J Neurosci 2007, 27: 12067–12077.PubMedCrossRefGoogle Scholar
  27. [27]
    Feick P, Haas SR, Singer MV, Böcker U. Low-dose exposure of intestinal epithelial cells to formaldehyde results in MAP kinase activation and molecular alteration of the focal adhesion protein paxillin. Toxicology 2006, 219: 60–72.PubMedCrossRefGoogle Scholar
  28. [28]
    Lim SK, Kim JC, Moon CJ, Kim GY, Han HJ, Park SH. Formaldehyde induces apoptosis through decreased Prx 2 via p38 MAPK in lung epithelial cells. Toxicology 2010, 271: 100–106.PubMedCrossRefGoogle Scholar
  29. [29]
    Medhurst SJ, Walker K, Bowes M, Kidd BL, Glatt M, Muller M, et al. A rat model of bone cancer pain. Pain 2002, 96: 129–140.PubMedCrossRefGoogle Scholar
  30. [30]
    Fujita M, Ueda T, Handa T. Generation of formaldehyde by pharmaceutical excipients and its absorption by meglumine. Chem Pharm Bull (Tokyo) 2009, 57: 1096–1099.CrossRefGoogle Scholar
  31. [31]
    O’Neil KA, Miller FR, Barder TJ, Lubman DM. Profiling the progression of cancer: separation of microsomal proteins in MCF10 breast epithelial cell lines using nonporous chromatophoresis. Proteomics 2003, 3: 1256–1269.PubMedCrossRefGoogle Scholar
  32. [32]
    Wang Y. The functional regulation of TRPV1 and its role in pain sensitization. Neurochem Res 2008, 33: 2008–2012.PubMedCrossRefGoogle Scholar
  33. [33]
    Cheng JK, Ji RR. Intracellular signaling in primary sensory neurons and persistent pain. Neurochem Res 2008, 33: 1970–1978.PubMedCrossRefGoogle Scholar
  34. [34]
    Bron R, Klesse LJ, Shah K, Parada LF, Winter J. Activation of Ras is necessary and sufficient for upregulation of vanilloid receptor type 1 in sensory neurons by neurotrophic factors. Mol Cell Neurosci 2003, 22: 118–132.PubMedCrossRefGoogle Scholar
  35. [35]
    Chen Y, Geis C, Sommer C. Activation of TRPV1 contributes to morphine tolerance: involvement of the mitogen-activated protein kinase signaling pathway. J Neurosci 2008, 28: 5836–5845.PubMedCrossRefGoogle Scholar
  36. [36]
    Pezet S, Marchand F, D’Mello R, Grist J, Clark AK, Malcangio M, et al. Phosphatidylinositol 3-kinase is a key mediator of central sensitization in painful inflammatory conditions. J Neurosci 2008, 28: 4261–4270.PubMedCrossRefGoogle Scholar
  37. [37]
    Bonnas C, Specht K, Spleiss O, Froehner S, Dietmann G, Krüger JM, et al. Effects of cold ischemia and inflammatory tumor microenvironment on detection of PI3K/AKT and MAPK pathway activation patterns in clinical cancer samples. Int J Cancer 2011 [Epub ahead of print]Google Scholar
  38. [38]
    Shi TJ, Huang P, Mulder J, Ceccatelli S, Hokfelt T. Expression of p-Akt in sensory neurons and spinal cord after peripheral nerve injury. Neurosignals 2009, 17: 203–212.PubMedCrossRefGoogle Scholar

Copyright information

© Shanghai Institutes for Biological Sciences, CAS and Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Ying Han
    • 1
  • Yan Li
    • 1
  • Xing Xiao
    • 1
  • Jia Liu
    • 1
  • Xiang-Ling Meng
    • 1
  • Feng-Yu Liu
    • 1
  • Guo-Gang Xing
    • 1
  • You Wan
    • 1
    • 2
    • 3
  1. 1.Neuroscience Research InstituteSchool of Basic Medical SciencesBeijingChina
  2. 2.Department of NeurobiologySchool of Basic Medical SciencesBeijingChina
  3. 3.Key Laboratory for Neuroscience, The Ministry of Education / The Ministry of HealthPeking UniversityBeijingChina

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