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Stabilizing and upregulating Axin with tankyrase inhibitor reverses 5-fluorouracil chemoresistance and proliferation by targeting the WNT/caveolin-1 axis in colorectal cancer cells

Abstract

Chemoresistance is a main obstacle for colorectal cancer treatment. In this study, we evaluated the effects and mechanisms of the WNT/β-catenin signaling pathway on the chemoresistance of SW480 and SW620 colorectal cancer cells. The activity of β-catenin was activated/inhibited by the small molecule compound GSK-3 inhibitor 6-bromo-indirubin-3’-oxime and the tankyrase inhibitor XAV939. The downstream target genes of the WNT/β-catenin signaling pathway were screened using a cDNA microarray and bioinformatics analysis. Apoptosis induced by 5-Fu, cell cycle distribution and expression levels of WNT/β-catenin/TCF12/caveolin-1 and multidrug resistance proteins were examed by flow cytometry and western blot after β-catenin activation/inhibition and caveolin-1 overexpression/interference. The effect and mechanism of XAV939 on proliferation and apoptosis induced by 5-Fu in xenograft tumors of nude mice were evaluated by immunohistochemistry and TUNEL staining. 6-Bromo-indirubin-3’-oxime treatment increased β-catenin expression by regulating GSK-3β phosphorylation, accompanied by upregulation of TCF12, caveolin-1, P-gp, and MRP2 and downregulation of apoptosis induced by 5-Fu. Conversely, XAV939 treatment decreased β-catenin expression by upregulating Axin, accompanied by downregulation of TCF12, Caveolin-1, P-gp, and MRP2 and upregulation of apoptosis induced by 5-Fu. The caveolin-1 gene was identified as an important downstream gene of the WNT/β-catenin signaling pathway. Caveolin-1 overexpression upregulated β-catenin expression, increased P-gp and MRP2 expression and decreased apoptosis induced by 5-Fu; conversely, caveolin-1 interference caused the opposite effects. In addition, in vivo experiments showed that XAV939 treatment reduced β-catenin expression, increased apoptosis induced by 5-Fu and repressed xenograft tumor growth. Our findings suggested that inhibition of WNT/β-catenin/TCF12/caveolin-1 provides a new promising therapeutic strategy for colorectal cancer treatment.

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Fig. 1: Functional annotations of the differentially expressed genes after treatment of SW480 and SW620 cells with BIO/XAV939 based on GenCLiP 2.0 online tool analysis.
Fig. 2: Expression levels of P-gp, MRP2, GSK-3β, p-GSK-3β, β-catenin, TCF-12, Cav-1 and Axin in SW480 and SW620 cells after treatment with BIO, XAV939 and 5-Fu for 24 h.
Fig. 3: Expression levels of TCF-12 and β-catenin in SW480 and SW620 cells after treatment with BIO/XAV939 or transfection with Si-TCF12 for 24 h.
Fig. 4: Expression levels of TCF12, β-catenin, P-gp, MRP2 and Cav-1 in SW480 and SW620 cells after transfection with Cav-1/Si-Cav-1 for 24 h.
Fig. 5: Effect of BIO, XAV939 and Cav-1 on 5-Fu-induced proliferation and apoptosis in SW480 and SW620 cells.
Fig. 6: Effects of BIO, XAV939 and Cav-1 on the cell cycle distribution of SW480 and SW620 cells.
Fig. 7: Effect and mechanism of XAV939 on chemotherapy in nude mice.
Fig. 8: BIO treatment regulate the translational activity of Cav-1 promoter.

Data availability

Data generated for the current study are available from the corresponding author on reasonable request.

References

  1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68:394–424.

    Article  Google Scholar 

  2. Giacchetti S, Itzhaki M, Gruia G, Adam R, Zidani R, Kunstlinger F, et al. Long-term survival of patients with unresectable colorectal cancer liver metastases following infusional chemotherapy with 5-fluorouracil, leucovorin, oxaliplatin and surgery. Ann Oncol. 1999;10:663–9.

    CAS  Article  Google Scholar 

  3. Shakibaei M, Kraehe P, Popper B, Shayan P, Goel A, Buhrmann C. Curcumin potentiates antitumor activity of 5-fluorouracil in a 3D alginate tumor microenvironment of colorectal cancer. BMC Cancer. 2015;15:250.

    Article  Google Scholar 

  4. Longley DB, Harkin DP, Johnston PG. 5-fluorouracil: Mechanisms of action and clinical strategies. Nat Rev Cancer. 2003;3:330–8.

    CAS  Article  Google Scholar 

  5. Bunz F, Hwang PM, Torrance C, Waldman T, Zhang Y, Dillehay L, et al. Disruption of p53 in human cancer cells alters the responses to therapeutic agents. J Clin Invest. 1999;104:263–9.

    CAS  Article  Google Scholar 

  6. Pugacheva EN, Ivanov AV, Kravchenko JE, Kopnin BP, Levine AJ, Chumakov PM. Novel gain of function activity of p53 mutants: Activation of the dUTPase gene expression leading to resistance to 5-fluorouracil. Oncogene 2002;21:4595–600.

    CAS  Article  Google Scholar 

  7. He L, Zhu H, Zhou S, Wu T, Wu H, Yang H, et al. Wnt pathway is involved in 5-FU drug resistance of colorectal cancer cells. Exp Mol Med. 2018;50:1–12.

    Article  Google Scholar 

  8. Yamada T, Takaoka AS, Naishiro Y, Hayashi R, Maruyama K, Maesawa C, et al. Transactivation of the multidrug resistance 1 gene by T-cell factor 4/beta-catenin complex in early colorectal carcinogenesis. Cancer Res. 2000;60:4761–6.

    CAS  PubMed  Google Scholar 

  9. Kioussi C, Briata P, Baek SH, Rose DW, Hamblet NS, Herman T, et al. Identification of a Wnt/Dvl/beta-Catenin —> Pitx2 pathway mediating cell-type-specific proliferation during development. Cell 2002;111:673–85.

    CAS  Article  Google Scholar 

  10. Tetsu O, McCormick F. Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature 1999;398:422–6.

    CAS  Article  Google Scholar 

  11. Liu K, Li J, Wu X, Chen M, Luo F, Li J. GSK-3β inhibitor 6-bromo-indirubin-3’-oxime promotes both adhesive activity and drug resistance in colorectal cancer cells. Int J Oncol. 2017;51:1821–30.

    CAS  Article  Google Scholar 

  12. Wu X, Luo F, Li J, Zhong X, Liu K. Tankyrase 1 inhibitior XAV939 increases chemosensitivity in colon cancer cell lines via inhibition of the Wnt signaling pathway. Int J Oncol. 2016;48:1333–40.

    CAS  Article  Google Scholar 

  13. Wang JH, Zhao LF, Lin P, Su XR, Chen SJ, Huang LQ, et al. GenCLiP 2.0: a web server for functional clustering of genes and construction of molecular networks based on free terms. Bioinformatics 2014;30:2534–6.

    CAS  Article  Google Scholar 

  14. Kent WJ, Sugnet CW, Furey TS, Roskin KM, Pringle TH, Zahler AM, et al. The human genome browser at UCSC. Genome Res. 2002;12:996–1006.

    CAS  Article  Google Scholar 

  15. Li VS, Ng SS, Boersema PJ, Low TY, Karthaus WR, Gerlach JP, et al. Wnt signaling through inhibition of β-catenin degradation in an intact Axin1 complex. Cell 2012;149:1245–56.

    CAS  Article  Google Scholar 

  16. Rubinfeld B, Albert I, Porfiri E, Fiol C, Munemitsu S, Polakis P. Binding of GSK3beta to the APC-beta-catenin complex and regulation of complex assembly. Science 1996;272:1023–6.

    CAS  Article  Google Scholar 

  17. Lepourcelet M, Chen YN, France DS, Wang H, Crews P, Petersen F, et al. Small-molecule antagonists of the oncogenic Tcf/beta-catenin protein complex. Cancer Cell. 2004;5:91–102.

    CAS  Article  Google Scholar 

  18. Sargiacomo M, Scherer PE, Tang Z, Kübler E, Song KS, Sanders MC, et al. Oligomeric structure of caveolin: implications for caveolae membrane organization. Proc Natl Acad Sci USA. 1995;92:9407–11.

    CAS  Article  Google Scholar 

  19. Rothberg KG, Heuser JE, Donzell WC, Ying YS, Glenney JR, Anderson RG. Caveolin, a protein component of caveolae membrane coats. Cell. 1992;68:673–82.

    CAS  Article  Google Scholar 

  20. Williams TM, Lisanti MP. The caveolin proteins. Genome Biol. 2004;5:214.

    Article  Google Scholar 

  21. Williams TM, Lisanti MP. Caveolin-1 in oncogenic transformation, cancer, and metastasis. Am J Physiol Cell Physiol. 2005;288:C494–506.

    CAS  Article  Google Scholar 

  22. Wang Z, Wang N, Liu P, Peng F, Tang H, Chen Q, et al. Caveolin-1, a stress-related oncotarget, in drug resistance. Oncotarget 2015;6:37135–50.

    Article  Google Scholar 

  23. Yang CP, Galbiati F, Volonte D, Horwitz SB, Lisanti MP. Upregulation of caveolin-1 and caveolae organelles in Taxol-resistant A549 cells. FEBS Lett. 1998;439:368–72.

    CAS  Article  Google Scholar 

  24. Lavie Y, Fiucci G, Liscovitch M. Up-regulation of caveolae and caveolar constituents in multidrug-resistant cancer cells. J Biol Chem. 1998;273:32380–3.

    CAS  Article  Google Scholar 

  25. Bélanger MM, Gaudreau M, Roussel E, Couet J. Role of caveolin-1 in etoposide resistance development in A549 lung cancer cells. Cancer Biol Ther. 2004;3:954–9.

    Article  Google Scholar 

  26. Li J, Hassan GS, Williams TM, Minetti C, Pestell RG, Tanowitz HB, et al. Loss of caveolin-1 causes the hyper-proliferation of intestinal crypt stem cells, with increased sensitivity to whole body gamma-radiation. Cell Cycle. 2005;4:1817–25.

    CAS  Article  Google Scholar 

  27. Tahir SA, Kurosaka S, Tanimoto R, Goltsov AA, Park S, Thompson TC. Serum caveolin-1, a biomarker of drug response and therapeutic target in prostate cancer models. Cancer Biol Ther. 2013;14:117–26.

    CAS  Article  Google Scholar 

  28. Yuan G, Regel I, Lian F, Friedrich T, Hitkova I, Hofheinz RD, et al. WNT6 is a novel target gene of caveolin-1 promoting chemoresistance to epirubicin in human gastric cancer cells. Oncogene 2013;32:375–87.

    CAS  Article  Google Scholar 

  29. Brodie SA, Lombardo C, Li G, Kowalski J, Gandhi K, You S, et al. Aberrant promoter methylation of caveolin-1 is associated with favorable response to taxane-platinum combination chemotherapy in advanced NSCLC. PLoS One. 2014;9:e107124.

    Article  Google Scholar 

  30. Cho YH, Ro EJ, Yoon JS, Mizutani T, Kang DW, Park JC, et al. 5-FU promotes stemness of colorectal cancer via p53-mediated WNT/β-catenin pathway activation. Nat Commun. 2020;11:5321.

    CAS  Article  Google Scholar 

  31. Jiang S, Miao D, Wang M, Lv J, Wang Y, Tong J. MiR-30-5p suppresses cell chemoresistance and stemness in colorectal cancer through USP22/Wnt/β-catenin signaling axis. J Cell Mol Med. 2019;23:630–40.

    CAS  Article  Google Scholar 

  32. Galbiati F, Volonte D, Brown AM, Weinstein DE, Ben-Ze’ev A, Pestell RG, et al. Caveolin-1 expression inhibits Wnt/beta-catenin/Lef-1 signaling by recruiting beta-catenin to caveolae membrane domains. J Biol Chem. 2000;275:23368–77.

    CAS  Article  Google Scholar 

  33. Yamamoto H, Komekado H, Kikuchi A. Caveolin is necessary for Wnt-3a-dependent internalization of LRP6 and accumulation of beta-catenin. Dev Cell. 2006;11:213–23.

    CAS  Article  Google Scholar 

  34. Arqués O, Chicote I, Puig I, Tenbaum SP, Argilés G, Dienstmann R, et al. Tankyrase Inhibition Blocks Wnt/β-Catenin Pathway and Reverts Resistance to PI3K and AKT Inhibitors in the Treatment of Colorectal Cancer. Clin Cancer Res. 2016;22:644–56.

    Article  Google Scholar 

  35. Waaler J, Machon O, Tumova L, Dinh H, Korinek V, Wilson SR, et al. A novel tankyrase inhibitor decreases canonical Wnt signaling in colon carcinoma cells and reduces tumor growth in conditional APC mutant mice. Cancer Res. 2012;72:2822–32.

    CAS  Article  Google Scholar 

  36. Lau T, Chan E, Callow M, Waaler J, Boggs J, Blake RA, et al. A novel tankyrase small-molecule inhibitor suppresses APC mutation-driven colorectal tumor growth. Cancer Res. 2013;73:3132–44.

    CAS  Article  Google Scholar 

  37. James RG, Davidson KC, Bosch KA, Biechele TL, Robin NC, Taylor RJ, et al. WIKI4, a novel inhibitor of tankyrase and Wnt/ß-catenin signaling. PLoS One. 2012;7:e50457.

    CAS  Article  Google Scholar 

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Funding

This study was sponsored by Guangdong Natural Science Foundation (no.2014A030307007, no.2017A030307005).

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L-KP, LF, and L-JB designed the experiments. LF and L-JH conducted the experiments. L-KP, LF and L-JB participated in the data analysis. L-KP, LF and L-JB drafted and polished the manuscript. All authors read and approved the final manuscript.

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Correspondence to Kunping Liu.

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All studies involving animals were performed following the National Guides for the Care and Use of Laboratory Animals and approved by the Institutional Animal Care and Use Committee of Guangzhou Medical University.

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Luo, F., Li, J., Liu, J. et al. Stabilizing and upregulating Axin with tankyrase inhibitor reverses 5-fluorouracil chemoresistance and proliferation by targeting the WNT/caveolin-1 axis in colorectal cancer cells. Cancer Gene Ther (2022). https://doi.org/10.1038/s41417-022-00493-y

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