Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

SphK2 confers 5-fluorouracil resistance to colorectal cancer via upregulating H3K56ac-mediated DPD expression


Aberrant sphingolipid metabolism has been implicated in chemoresistance, but the underlying mechanisms are still poorly understood. Herein we revealed a previously unrecognized mechanism of 5-fluorouracil (5-FU) resistance contributed by high SphK2-upregulated dihydropyrimidine dehydrogenase (DPD) in colorectal cancer (CRC), which is evidenced from human CRC specimens, animal models, and cancer cell lines. TMA samples from randomly selected 60 CRC specimens firstly identified the clinical correlation between high SphK2 and increased DPD (p < 0.001). Then the regulatory mechanism was explored in CRC models of villin-SphK2 Tg mice, SphK2−/mice, and human CRC cells xenografted nude mice. Assays of ChIP-Seq and luciferase reporter gene demonstrated that high SphK2 upregulated DPD through promoting the HDAC1-mediated H3K56ac, leading to the degradation of intracellular 5-FU into inactive α-fluoro-β-alanine (FBAL). Lastly, inhibition of SphK2 by SLR080811 exhibited excellent inhibition on DPD expression and potently reversed 5-FU resistance in colorectal tumors of villin-SphK2 Tg mice. Overall, this study manifests that SphK2high conferred 5-FU resistance through upregulating tumoral DPD, which highlights the strategies of blocking SphK2 to overcome 5-FU resistance in CRC.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: High SphK2 but not SphK1 correlates with increased DPD in human CRC specimens.
Fig. 2: SphK2 Tg mice demonstrate resistance to 5-FU therapy due to their high expressions of tumoral DPD.
Fig. 3: HCT116sphk2 cells resisted 5-FU treatment due to increased degradation of the intracellular 5-FU.
Fig. 4: SphK2-mediated 5-FU resistance attributed to increased DPD in HCT116sphk2 cells xenografted nude mice.
Fig. 5: SphK2 upregulates DPYD expression through promoting the acylation of H3K56.
Fig. 6: Inhibition of SphK2 by SLR080811 effectively reversed 5-FU resistance by downregulating the expression of tumoral DPD.


  1. 1.

    Van Cutsem E, Twelves C, Cassidy J, Allman D, Bajetta E, Boyer M, et al. Oral capecitabine compared with intravenous fluorouracil plus leucovorin in patients with metastatic colorectal cancer: results of a large phase III study. J Clin Oncol. 2001;19:4097–106.

    Article  Google Scholar 

  2. 2.

    Manca A, Asseburg C, Bravo VY, Seymour MT, Meade A, Stephens R, et al. The cost-effectiveness of different chemotherapy strategies for patients with poor prognosis advanced colorectal cancer (MRC FOCUS). Value Health. 2012;15:22–31.

    Article  Google Scholar 

  3. 3.

    Seymour MT, Maughan TS, Ledermann JA, Topham C, James R, Gwyther SJ, et al. Different strategies of sequential and combination chemotherapy for patients with poor prognosis advanced colorectal cancer (MRC FOCUS): a randomised controlled trial. Lancet. 2007;370:143–52.

    CAS  Article  Google Scholar 

  4. 4.

    Afzal S, Gusella M, Vainer B, Vogel UB, Andersen JT, Broedbaek K, et al. Combinations of polymorphisms in genes involved in the 5-Fluorouracil metabolism pathway are associated with gastrointestinal toxicity in chemotherapy-treated colorectal cancer patients. Clin Cancer Res. 2011;17:3822–9.

    CAS  Article  Google Scholar 

  5. 5.

    Peters GJ, Backus HH, Freemantle S, van Triest B, Codacci-Pisanelli G, van der Wilt CL, et al. Induction of thymidylate synthase as a 5-fluorouracil resistance mechanism. Biochim Biophys Acta. 2002;1587:194–205.

    CAS  Article  Google Scholar 

  6. 6.

    Hamajima N, Matsuda K, Sakata S, Tamaki N, Sasaki M, Nonaka M. A novel gene family defined by human dihydropyrimidinase and three related proteins with differential tissue distribution. Gene. 1996;180:157–63.

    CAS  Article  Google Scholar 

  7. 7.

    Offer SM, Wegner NJ, Fossum C, Wang K, Diasio RB. Phenotypic profiling of DPYD variations relevant to 5-fluorouracil sensitivity using real-time cellular analysis and in vitro measurement of enzyme activity. Cancer Res. 2013;73:1958–68.

    CAS  Article  Google Scholar 

  8. 8.

    Lee AM, Shi Q, Pavey E, Alberts SR, Sargent DJ, Sinicrope FA, et al. DPYD variants as predictors of 5-fluorouracil toxicity in adjuvant colon cancer treatment (NCCTG N0147). J Natl Cancer Inst. 2014;106:dju298.

    PubMed  PubMed Central  Google Scholar 

  9. 9.

    Spiegel S, Milstien S. Sphingosine-1-phosphate: an enigmatic signalling lipid. Nat Rev Mol Cell Biol. 2003;4:397–407.

    CAS  Article  Google Scholar 

  10. 10.

    Min J, Traynor D, Stegner AL, Zhang L, Hanigan MH, Alexander H, et al. Sphingosine kinase regulates the sensitivity of Dictyostelium discoideum cells to the anticancer drug cisplatin. Eukaryot Cell. 2005;4:178–89.

    CAS  Article  Google Scholar 

  11. 11.

    Salas A, Ponnusamy S, Senkal CE, Meyers-Needham M, Selvam SP, Saddoughi SA, et al. Sphingosine kinase-1 and sphingosine 1-phosphate receptor 2 mediate Bcr-Abl1 stability and drug resistance by modulation of protein phosphatase 2A. Blood. 2011;117:5941–52.

    CAS  Article  Google Scholar 

  12. 12.

    Rosa R, Marciano R, Malapelle U, Formisano L, Nappi L, D’Amato C, et al. Sphingosine kinase 1 overexpression contributes to cetuximab resistance in human colorectal cancer models. Clin Cancer Res. 2013;19:138–47.

    CAS  Article  Google Scholar 

  13. 13.

    Hait NC, Allegood J, Maceyka M, Strub GM, Harikumar KB, Singh SK, et al. Regulation of histone acetylation in the nucleus by sphingosine-1-phosphate. Science. 2009;325:1254–7.

    CAS  Article  Google Scholar 

  14. 14.

    Grassi S, Mauri L, Prioni S, Cabitta L, Sonnino S, Prinetti A, et al. Sphingosine 1-phosphate receptors and metabolic enzymes as druggable targets for brain diseases. Front Pharm. 2019;10:807.

    CAS  Article  Google Scholar 

  15. 15.

    O’Sullivan S, Dev KK. Sphingosine-1-phosphate receptor therapies: advances in clinical trials for CNS-related diseases. Neuropharmacology. 2017;113:597–607.

    Article  Google Scholar 

  16. 16.

    French KJ, Zhuang Y, Maines LW, Gao P, Wang W, Beljanski V, et al. Pharmacology and antitumor activity of ABC294640, a selective inhibitor of sphingosine kinase-2. J Pharm Exp Ther. 2010;333:129–39.

    CAS  Article  Google Scholar 

  17. 17.

    Fitzpatrick LR, Green C, Frauenhoffer EE, French KJ, Zhuang Y, Maines LW, et al. Attenuation of arthritis in rodents by a novel orally-available inhibitor of sphingosine kinase. Inflammopharmacology. 2011;19:75–87.

    CAS  Article  Google Scholar 

  18. 18.

    Venant H, Rahmaniyan M, Jones EE, Lu P, Lilly MB, Garrett-Mayer E, et al. The sphingosine kinase 2 inhibitor ABC294640 reduces the growth of prostate cancer cells and results in accumulation of dihydroceramides in vitro and in vivo. Mol Cancer Ther. 2015;14:2744–52.

    CAS  Article  Google Scholar 

  19. 19.

    Britten CD, Garrett-Mayer E, Chin SH, Shirai K, Ogretmen B, Bentz TA, et al. A Phase I Study of ABC294640, a first-in-class sphingosine kinase-2 inhibitor, in patients with advanced solid tumors. Clin Cancer Res. 2017;23:4642–50.

    CAS  Article  Google Scholar 

  20. 20.

    Kharel Y, Raje M, Gao M, Gellett AM, Tomsig JL, Lynch KR, et al. Sphingosine kinase type 2 inhibition elevates circulating sphingosine 1-phosphate. Biochem J. 2012;447:149–57.

    CAS  Article  Google Scholar 

  21. 21.

    Liang J, Nagahashi M, Kim EY, Harikumar KB, Yamada A, Huang WC, et al. Sphingosine-1-phosphate links persistent STAT3 activation, chronic intestinal inflammation, and development of colitis-associated cancer. Cancer Cell. 2013;23:107–20.

    CAS  Article  Google Scholar 

  22. 22.

    Adjei AA, Reid JM, Diasio RB, Sloan JA, Smith DA, Rubin J, et al. Comparative pharmacokinetic study of continuous venous infusion fluorouracil and oral fluorouracil with eniluracil in patients with advanced solid tumors. J Clin Oncol. 2002;20:1683–91.

    CAS  Article  Google Scholar 

  23. 23.

    Micoli G, Turci R, Arpellini M, Minoia C. Determination of 5-fluorouracil in environmental samples by solid-phase extraction and high-performance liquid chromatography with ultraviolet detection. J Chromatogr B Biomed Sci Appl. 2001;750:25–32.

    CAS  Article  Google Scholar 

  24. 24.

    Dovey OM, Foster CT, Cowley SM. Histone deacetylase 1 (HDAC1), but not HDAC2, controls embryonic stem cell differentiation. Proc Natl Acad Sci USA. 2010;107:8242–7.

    CAS  Article  Google Scholar 

  25. 25.

    Maceyka M, Harikumar KB, Milstien S, Spiegel S. Sphingosine-1-phosphate signaling and its role in disease. Trends Cell Biol. 2012;22:50–60.

    CAS  Article  Google Scholar 

  26. 26.

    Guillermet-Guibert J, Davenne L, Pchejetski D, Saint-Laurent N, Brizuela L, Guilbeau-Frugier C, et al. Targeting the sphingolipid metabolism to defeat pancreatic cancer cell resistance to the chemotherapeutic gemcitabine drug. Mol Cancer Ther. 2009;8:809–20.

    CAS  Article  Google Scholar 

  27. 27.

    Sarkar S, Maceyka M, Hait NC, Paugh SW, Sankala H, Milstien S, et al. Sphingosine kinase 1 is required for migration, proliferation and survival of MCF-7 human breast cancer cells. Febs Lett. 2005;579:5313–7.

    CAS  Article  Google Scholar 

  28. 28.

    Rosmarin D, Palles C, Pagnamenta A, Kaur K, Pita G, Martin M, et al. A candidate gene study of capecitabine-related toxicity in colorectal cancer identifies new toxicity variants at DPYD and a putative role for ENOSF1 rather than TYMS. Gut. 2015;64:111–20.

    CAS  Article  Google Scholar 

  29. 29.

    Amstutz U, Henricks LM, Offer SM, Barbarino J, Schellens J, Swen JJ, et al. Clinical pharmacogenetics implementation consortium (CPIC) guideline for dihydropyrimidine dehydrogenase genotype and fluoropyrimidine dosing: 2017 update. Clin Pharm Ther. 2018;103:210–6.

    CAS  Article  Google Scholar 

  30. 30.

    Xun C, Chen MB, Qi L, Tie-Ning Z, Peng X, Ning L, et al. Targeting sphingosine kinase 2 (SphK2) by ABC294640 inhibits colorectal cancer cell growth in vitro and in vivo. J Exp Clin Cancer Res. 2015;34:94.

    Article  Google Scholar 

  31. 31.

    Camp ER, Patterson LD, Kester M, Voelkel-Johnson C. Therapeutic implications of bioactive sphingolipids: a focus on colorectal cancer. Cancer Biol Ther. 2017;18:640–50.

    CAS  Article  Google Scholar 

  32. 32.

    Parang B, Barrett CW, Williams CS. AOM/DSS model of colitis-associated cancer. Methods Mol Biol. 2016;1422:297–307.

    CAS  Article  Google Scholar 

  33. 33.

    Sun C, Zargham R, Shao Q, Gui X, Marcus V, Lazaris A, et al. Association of CD98, integrin beta1, integrin beta3 and Fak with the progression and liver metastases of colorectal cancer. Pathol Res Pr. 2014;210:668–74.

    CAS  Article  Google Scholar 

  34. 34.

    Kidder BL, Hu G, Zhao K. ChIP-Seq: technical considerations for obtaining high-quality data. Nat Immunol. 2011;12:918–22.

    CAS  Article  Google Scholar 

Download references


We thank Prof Zu-Hua Gao at Department of Pathology, McGill University, for helpful discussions and data analysis.


This work was supported by National Natural Science Foundation of China (91629303/81673449/81872884/81973350) and Beijing Natural Science Foundation and Scientific Research Program of Municipal Commission of Education (KZ201710025020/KZ201810025033).

Author information




XJQ conceived the project. YHZ performed the experiments of molecular mechanisms. WNS and SHW provided clinical samples and pathological analysis. RRM and SYS conducted animal studies. DDL and SBW implemented experiments of pharmaceutical analysis. SXC, ZKG, and WYW performed statistical analysis. YHZ wrote the paper, which was edited by all authors.

Corresponding authors

Correspondence to Shu-Xiang Cui or Xian-Jun Qu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zhang, YH., Shi, WN., Wu, SH. et al. SphK2 confers 5-fluorouracil resistance to colorectal cancer via upregulating H3K56ac-mediated DPD expression. Oncogene 39, 5214–5227 (2020).

Download citation

Further reading


Quick links