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

Abstract

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.

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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.

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Acknowledgements

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

Funding

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).

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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.

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

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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). https://doi.org/10.1038/s41388-020-1352-y

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