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5-Fluorouracil inhibits cell migration by induction of Sestrin2 in colon cancer cells

Archives of Pharmacal Research volume 40, pages 231–239 (2017)Cite this article

Abstract

5-Fluorouracil (5-FU) is a chemotherapeutic agent used in the treatment of colorectal cancer. In this study, we investigated whether 5-FU induces Sestrin2 (SESN2), an antioxidant enzyme, and the role of SESN2 in 5-FU action in colon cancer cells. We found that 5-FU upregulated SESN2 protein expression in both HCT116 and HT29 cells. It also increased transcripts of SESN1 and SESN2, but not of SESN3. Furthermore, we investigated whether production of reactive oxygen species (ROS) was involved in 5-FU-induced SESN2 expression. 5-FU did not increase ROS production nor affect Nrf2 phosphorylation and expression levels. Moreover, SESN2 upregulation by 5-FU was not prevented by pretreatment with antioxidants. Next, we investigated p53 levels after 5-FU treatment to elucidate the regulation of SESN2 by 5-FU. An increase in p53 levels was detected following 5-FU treatment; pifithrin-α, an inhibitor of p53 activation, reversed 5-FU-induced SESN2 expression. 5-FU prevented serum-induced in vitro cell migration, but knockdown of SESN2 or treatment with pifithrin-α reversed a 5-FU-mediated decrease in cell migration. Taken together, our results suggest that 5-FU increases SESN2 levels via a p53-dependent pathway, which contributes to inhibition of cancer cell migration in vitro.

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References

  • Buckbinder L, Talbott R, Seizinger BR, Kley N (1994) Gene regulation by temperature-sensitive p53 mutants: identification of p53 response genes. Proc Natl Acad Sci USA 91:10640–10644

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Budanov AV (2014) The role of tumor suppressor p53 in the antioxidant defense and metabolism. Subcell Biochem 85:337–358

    Article  PubMed  PubMed Central  Google Scholar 

  • Budanov AV, Karin M (2008) p53 target genes Sestrin1 and Sestrin2 connect genotoxic stress and mTOR signaling. Cell 134:451–460

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Budanov AV, Shoshani T, Faerman A, Zelin E, Kamer I, Kalinski H, Gorodin S, Fishman A, Chajut A, Einat P, Skaliter R, Gudkov AV, Chumakov PM, Feinstein E (2002) Identification of a novel stress-responsive gene Hi95 involved in regulation of cell viability. Oncogene 21:6017–6031

    CAS  Article  PubMed  Google Scholar 

  • Budanov AV, Sablina AA, Feinstein E, Koonin EV, Chumakov PM (2004) Regeneration of peroxiredoxins by p53-regulated sestrins, homologs of bacterial AhpD. Science 304:596–600

    CAS  Article  PubMed  Google Scholar 

  • Budanov AV, Lee JH, Karin M (2010) Stressin’ Sestrins take an aging fight. EMBO Mol Med 2:388–400

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Cairns RA, Harris IS, Mak TW (2011) Regulation of cancer cell metabolism. Nat Rev Cancer 11:85–95

    CAS  Article  PubMed  Google Scholar 

  • Cannito S, Novo E, Compagnone A, Valfre di Bonzo L, Busletta C, Zamara E, Paternostro C, Povero D, Bandino A, Bozzo F, Cravanzola C, Bravoco V, Colombatto S, Parola M (2008) Redox mechanisms switch on hypoxia-dependent epithelial-mesenchymal transition in cancer cells. Carcinogenesis 29:2267–2278

    CAS  Article  PubMed  Google Scholar 

  • Chen CC, Jeon SM, Bhaskar PT, Nogueira V, Sundararajan D, Tonic I, Park Y, Hay N (2010) FoxOs inhibit mTORC1 and activate Akt by inducing the expression of Sestrin3 and Rictor. Dev Cell 18:592–604

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Chen KB, Xuan Y, Shi WJ, Chi F, Xing R, Zeng YC (2016) Sestrin2 expression is a favorable prognostic factor in patients with non-small cell lung cancer. Am J Transl Res 8:1903–1909

    PubMed  PubMed Central  Google Scholar 

  • Ferrando M, Gueron G, Elguero B, Giudice J, Salles A, Leskow FC, Jares-Erijman EA, Colombo L, Meiss R, Navone N, De Siervi A, Vazquez E (2011) Heme oxygenase 1 (HO-1) challenges the angiogenic switch in prostate cancer. Angiogenesis 14:467–479

    CAS  Article  PubMed  Google Scholar 

  • Galanis A, Pappa A, Giannakakis A, Lanitis E, Dangaj D, Sandaltzopoulos R (2008) Reactive oxygen species and HIF-1 signalling in cancer. Cancer Lett 266:12–20

    CAS  Article  PubMed  Google Scholar 

  • Giatromanolaki A, Sivridis E, Koukourakis MI (2006) Angiogenesis in colorectal cancer: prognostic and therapeutic implications. Am J Clin Oncol 29:408–417

    Article  PubMed  Google Scholar 

  • Guina T, Biasi F, Calfapietra S, Nano M, Poli G (2015) Inflammatory and redox reactions in colorectal carcinogenesis. Ann N.Y. Acad Sci 1340:95–103

    CAS  Article  PubMed  Google Scholar 

  • Haggar FA, Boushey RP (2009) Colorectal cancer epidemiology: incidence, mortality, survival, and risk factors. Clin Colon Rectal Surg 22:191–197

    Article  PubMed  PubMed Central  Google Scholar 

  • Hill M, Pereira V, Chauveau C, Zagani R, Remy S, Tesson L, Mazal D, Ubillos L, Brion R, Asghar K, Mashreghi MF, Kotsch K, Moffett J, Doebis C, Seifert M, Boczkowski J, Osinaga E, Anegon I (2005) Heme oxygenase-1 inhibits rat and human breast cancer cell proliferation: mutual cross inhibition with indoleamine 2,3-dioxygenase. FASEB J 19:1957–1968

    CAS  Article  PubMed  Google Scholar 

  • Hwang PM, Bunz F, Yu J, Rago C, Chan TA, Murphy MP, Kelso GF, Smith RA, Kinzler KW, Vogelstein B (2001) Ferredoxin reductase affects p53-dependent, 5-fluorouracil-induced apoptosis in colorectal cancer cells. Nat Med 7:1111–1117

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Komarov PG, Komarova EA, Kondratov RV, Christov-Tselkov K, Coon JS, Chernov MV, Gudkov AV (1999) A chemical inhibitor of p53 that protects mice from the side effects of cancer therapy. Science 285:1733–1737

    CAS  Article  PubMed  Google Scholar 

  • Lee WH, Kim YW, Choi JH, Brooks SC 3rd, Lee MO, Kim SG (2009) Oltipraz and dithiolethione congeners inhibit hypoxia-inducible factor-1α activity through p70 ribosomal S6 kinase-1α inhibition and H2O2-scavenging effect. Mol Cancer Ther 8:2791–2802

    CAS  Article  PubMed  Google Scholar 

  • Lee JH, Budanov AV, Karin M (2013) Sestrins orchestrate cellular metabolism to attenuate aging. Cell Metab 18:792–801

    CAS  Article  PubMed  Google Scholar 

  • Longley DB, Harkin DP, Johnston PG (2003) 5-Fluorouracil: mechanisms of action and clinical strategies. Nat Rev Cancer 3:330–338

    CAS  Article  PubMed  Google Scholar 

  • Niture SK, Kaspar JW, Shen J, Jaiswal AK (2010) Nrf2 signaling and cell survival. Toxicol Appl Pharmacol 244:37–42

    CAS  Article  PubMed  Google Scholar 

  • Oh CM, Won YJ, Jung KW, Kong HJ, Cho H, Lee JK, Lee DH, Lee KH (2016) Cancer statistics in Korea: incidence, mortality, survival, and prevalence in 2013. Cancer Res Treat 48:436–450

    Article  PubMed  PubMed Central  Google Scholar 

  • Ro SH, Xue X, Ramakrishnan SK, Cho CS, Namkoong S, Jang I, Semple IA, Ho A, Park HW, Shah YM, Lee JH (2016) Tumor suppressive role of Sestrin2 during colitis and colon carcinogenesis. Elife 5:e12204

    PubMed  PubMed Central  Google Scholar 

  • Sanli T, Linher-Melville K, Tsakiridis T, Singh G (2012) Sestrin2 modulates AMPK subunit expression and its response to ionizing radiation in breast cancer cells. PLoS ONE 7:e32035

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Shimojo Y, Akimoto M, Hisanaga T, Tanaka T, Tajima Y, Honma Y, Takenaga K (2013) Attenuation of reactive oxygen species by antioxidants suppresses hypoxia-induced epithelial-mesenchymal transition and metastasis of pancreatic cancer cells. Clin Exp Metastasis 30:143–154

    CAS  Article  PubMed  Google Scholar 

  • Shin BY, Jin SH, Cho IJ, Ki SH (2012) Nrf2-ARE pathway regulates induction of Sestrin-2 expression. Free Radic Biol Med 53:834–841

    CAS  Article  PubMed  Google Scholar 

  • Takano H, Momota Y, Kani K, Aota K, Yamamura Y, Yamanoi T, Azuma M (2015) γ-Tocotrienol prevents 5-FU-induced reactive oxygen species production in human oral keratinocytes through the stabilization of 5-FU-induced activation of Nrf2. Int J Oncol 46:1453–1460

    CAS  PubMed  PubMed Central  Google Scholar 

  • Tomioka H, Mukohara T, Kataoka Y, Ekyalongo RC, Funakoshi Y, Imai Y, Kiyota N, Fujiwara Y, Minami H (2012) Inhibition of the mTOR/S6 K signal is necessary to enhance fluorouracil-induced apoptosis in gastric cancer cells with HER2 amplification. Int J Oncol 41:551–558

    CAS  PubMed  Google Scholar 

  • Velasco-Miguel S, Buckbinder L, Jean P, Gelbert L, Talbott R, Laidlaw J, Seizinger B, Kley N (1999) PA26, a novel target of the p53 tumor suppressor and member of the GADD family of DNA damage and growth arrest inducible genes. Oncogene 18:127–137

    CAS  Article  PubMed  Google Scholar 

  • Wang N, Pan W, Zhu M, Zhang M, Hao X, Liang G, Feng Y (2011) Fangchinoline induces autophagic cell death via p53/Sestrin2/AMPK signalling in human hepatocellular carcinoma cells. Br J Pharmacol 164:731–742

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Wang H, Zhang L, Yang X, Jin Y, Pei S, Zhang D, Zhang H, Zhou B, Zhang Y, Lin D (2015) PUMA mediates the combinational therapy of 5-FU and NVP-BEZ235 in colon cancer. Oncotarget 6:14385–14398

    Article  PubMed  PubMed Central  Google Scholar 

  • Wei JL, Fu ZX, Fang M, Guo JB, Zhao QN, Lu WD, Zhou QY (2015) Decreased expression of sestrin 2 predicts unfavorable outcome in colorectal cancer. Oncol Rep 33:1349–1357

    CAS  PubMed  Google Scholar 

  • Wolpin BM, Mayer RJ (2008) Systemic treatment of colorectal cancer. Gastroenterology 134:1296–1310

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Xu H, Sun H, Zhang H, Liu J, Fan F, Li Y, Ning X, Sun Y, Dai S, Liu B, Gao M, Fu S, Zhou C (2015) An shRNA based genetic screen identified Sesn2 as a potential tumor suppressor in lung cancer via suppression of Akt-mTOR-p70S6 K signaling. PLoS ONE 10:e0124033

    Article  PubMed  PubMed Central  Google Scholar 

  • Yasuda M, Kato S, Yamanaka N, Iimori M, Utsumi D, Kitahara Y, Iwata K, Matsuno K, Amagase K, Yabe-Nishimura C, Takeuchi K (2012) Potential role of the NADPH oxidase NOX1 in the pathogenesis of 5-fluorouracil-induced intestinal mucositis in mice. Am J Physiol Gastrointest Liver Physiol 302:G1133–G1142

    CAS  Article  PubMed  Google Scholar 

  • Zamora-Ros R, Fedirko V, Trichopoulou A, Gonzalez CA, Bamia C, Trepo E, Nothlings U, Duarte-Salles T, Serafini M, Bredsdorff L, Overvad K, Tjonneland A, Halkjaer J, Fagherazzi G, Perquier F, Boutron-Ruault MC, Katzke V, Lukanova A, Floegel A, Boeing H, Lagiou P, Trichopoulos D, Saieva C, Agnoli C, Mattiello A, Tumino R, Sacerdote C, Bueno-de-Mesquita HB, Peeters PH, Weiderpass E, Engeset D, Skeie G, Arguelles MV, Molina-Montes E, Dorronsoro M, Tormo MJ, Ardanaz E, Ericson U, Sonestedt E, Sund M, Landberg R, Khaw KT, Wareham NJ, Crowe FL, Riboli E, Jenab M (2013a) Dietary flavonoid, lignan and antioxidant capacity and risk of hepatocellular carcinoma in the European prospective investigation into cancer and nutrition study. Int J Cancer 133:2429–2443

    CAS  Article  PubMed  Google Scholar 

  • Zamora-Ros R, Not C, Guino E, Lujan-Barroso L, Garcia RM, Biondo S, Salazar R, Moreno V (2013b) Association between habitual dietary flavonoid and lignan intake and colorectal cancer in a Spanish case-control study (the Bellvitge colorectal cancer study). Cancer Causes Control 24:549–557

    Article  PubMed  Google Scholar 

  • Zhou Y, Zheng J, Li Y, Xu DP, Li S, Chen YM, Li HB (2016) Natural polyphenols for prevention and treatment of cancer. Nutrients 8(8):515

    Article  PubMed Central  Google Scholar 

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Acknowledgement

This study was supported by a research fund from Chosun University (2013).

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Affiliations

  1. College of Pharmacy, Chosun University, 309 Pilmun-daero, Dong-gu, Gwangju, 501-759, South Korea

    Kyuhwa Seo, Sung Hwan Ki & Sang Mi Shin

  2. College of Pharmacy, Mokpo National University, Muan-gun, Jeonnam, 534-729, South Korea

    Eun Young Park

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  1. Kyuhwa Seo
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  2. Sung Hwan Ki
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Correspondence to Sang Mi Shin.

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Seo, K., Ki, S.H., Park, E.Y. et al. 5-Fluorouracil inhibits cell migration by induction of Sestrin2 in colon cancer cells. Arch. Pharm. Res. 40, 231–239 (2017). https://doi.org/10.1007/s12272-016-0878-6

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  • DOI: https://doi.org/10.1007/s12272-016-0878-6

Keywords

  • 5-Fluorouracil
  • Sestrin2
  • p53
  • Cell migration
  • Colorectal cancer