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Original Paper

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Reduction of NANOG Mediates the Inhibitory Effect of Aspirin on Tumor Growth and Stemness in Colorectal Cancer

Wang H.a,b,c · Liu B.a · Wang J.a,d · Li J.e · Gong Y.f · Li S.a · Wang C.a · Cui B.a · Xue X.a · Yang M.a · Fan W.a · Kang Z.g · Kamran M.a · Xu J.a · Tian P.h · Luo Y.a · Hou Z.a · Dong L.a · Ren Y.h · Li M.f · Wen Q.e · Cheng W.a · Xu L.f · Wang L.h · Liu Q.a,b

Author affiliations

aInstitute of Cancer Stem Cell, Cancer Center, Dalian Medical University, Dalian, China
bState Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University, Guangzhou, China
cDepartment of Biochemistry and Molecular Biology, Harbin Medical University, Harbin, China
dDepartment of Oncology, The First Affiliated Hospital of Gannan Medical College, Ganzhou, China
eDepartment of Anesthesia, The First Affiliated Hospital, Dalian Medical University, Dalian, China
fDepartment of Oncology, The Second Affiliated Hospital, Dalian Medical University, Dalian, China
gDepartment of Hematology, The Second Affiliated Hospital, Dalian Medical University, Dalian, China
hDepartment of Oncology, The First Affiliated Hospital, Dalian Medical University, Dalian, China

Corresponding Author

Q Liu, L Wang, LZ Xu and W Cheng

Institute Cancer Stem Cell, Dalian Medical University, Dept Oncol, The First Affiliated Hospital,

Dalian Medical University, Dept Oncol, The Second Affiliated Hospital,

Dalian Medical University, Institute Cancer Stem Cell, Dalian Medical University (China);

E-Mail liuq9@mail.sysu.edu.cn/Whwl@hotmail.com/xvlingzhi@sina.com/wcheng@dmu.edu.cn

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Cell Physiol Biochem 2017;44:1051–1063

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Background/Aims: Cancer stem cells (CSCs) are considered to be responsible for tumor relapse and metastasis, which serve as a potential therapeutic target for cancer. Aspirin has been shown to reduce cancer risk and mortality, particularly in colorectal cancer. However, the CSCs-suppressing effect of aspirin and its relevant mechanisms in colorectal cancer remain unclear. Methods: CCK8 assay was employed to detect the cell viability. Sphere formation assay, colony formation assay, and ALDH1 assay were performed to identify the effects of aspirin on CSC properties. Western blotting was performed to detect the expression of the stemness factors. Xenograft model was employed to identify the anti-cancer effects of aspirin in vivo. Unpaired Student t test, ANOVA test and Kruskal-Wallis test were used for the statistical comparisons. Results: Aspirin attenuated colonosphere formation and decreased the ALDH1 positive cell population of colorectal cancer cells. Aspirin inhibited xenograft tumor growth and reduced tumor cells stemness in nude mice. Consistently, aspirin decreased the protein expression of stemness-related transcription factors, including c-Myc, OCT4 and NANOG. Suppression of NANOG blocked the effect of aspirin on sphere formation. Conversely, ectopic expression of NANOG rescued the aspirin-repressed sphere formation, suggesting that NANOG is a key downstream target. Moreover, we found that aspirin repressed NANOG expression in protein level by decreasing its stability. Conclusion: We have provided new evidence that aspirin attenuates CSC properties through down-regulation of NANOG, suggesting aspirin as a promising therapeutic agent for colorectal cancer treatment.

© 2017 The Author(s). Published by S. Karger AG, Basel


  1. WHO: World Cancer Report 2014, 2014.
  2. Nagtegaal ID, Quirke P: What is the role for the circumferential margin in the modern treatment of rectal cancer? J Clin Oncol 2008; 26: 303-312.
  3. Papadimitriou CA, Papakostas P, Karina M, Malettou L, Dimopoulos MA, Pentheroudakis G, Samantas E, Bamias A, Miliaras D, Basdanis G, Xiros N, Klouvas G, Bafaloukos D, Kafiri G, Papaspirou I, Pectasides D, Karanikiotis C, Economopoulos T, Efstratiou I, Korantzis I, Pisanidis N, Makatsoris T, Matsiakou F, Aravantinos G, Kalofonos HP, Fountzilas G: A randomized phase III trial of adjuvant chemotherapy with irinotecan, leucovorin and fluorouracil versus leucovorin and fluorouracil for stage II and III colon cancer: a Hellenic Cooperative Oncology Group study. BMC Med 2011; 9: 10.
  4. Gray R, Barnwell J, McConkey C, Hills RK, Williams NS, Kerr DJ: Adjuvant chemotherapy versus observation in patients with colorectal cancer: a randomised study. Lancet 2007; 370: 2020-2029.
  5. O’Brien CA, Pollett A, Gallinger S, Dick JE: A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 2007; 445: 106-110.
  6. Colak S, Zimberlin CD, Fessler E, Hogdal L, Prasetyanti PR, Grandela CM, Letai A, Medema JP: Decreased mitochondrial priming determines chemoresistance of colon cancer stem cells. Cell Death Differ 2014; 21: 1170-1177.
  7. Dylla SJ, Beviglia L, Park IK, Chartier C, Raval J, Ngan L, Pickell K, Aguilar J, Lazetic S, Smith-Berdan S, Clarke MF, Hoey T, Lewicki J, Gurney AL: Colorectal cancer stem cells are enriched in xenogeneic tumors following chemotherapy. PLoS One 2008; 3:e2428.
  8. Ricci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C, De Maria R: Identification and expansion of human colon-cancer-initiating cells. Nature 2007; 445: 111-115.
  9. Kemper K, Prasetyanti PR, De Lau W, Rodermond H, Clevers H, Medema JP: Monoclonal antibodies against Lgr5 identify human colorectal cancer stem cells. Stem Cells 2012; 30: 2378-2386.
  10. Jung P, Sato T, Merlos-Suarez A, Barriga FM, Iglesias M, Rossell D, Auer H, Gallardo M, Blasco MA, Sancho E, Clevers H, Batlle E: Isolation and in vitro expansion of human colonic stem cells. Nat Med 2011; 17: 1225-1227.
  11. Todaro M, Gaggianesi M, Catalano V, Benfante A, Iovino F, Biffoni M, Apuzzo T, Sperduti I, Volpe S, Cocorullo G, Gulotta G, Dieli F, De Maria R, Stassi G: CD44v6 is a marker of constitutive and reprogrammed cancer stem cells driving colon cancer metastasis. Cell Stem Cell 2014; 14: 342-356.
  12. Huang EH, Hynes MJ, Zhang T, Ginestier C, Dontu G, Appelman H, Fields JZ, Wicha MS, Boman BM: Aldehyde dehydrogenase 1 is a marker for normal and malignant human colonic stem cells (SC) and tracks SC overpopulation during colon tumorigenesis. Cancer Res 2009; 69: 3382-3389.
  13. Tica Sedlar I, Petricevic J, Saraga-Babic M, Pintaric I, Vukojevic K: Apoptotic pathways and stemness in the colorectal epithelium and lamina propria mucosae during the human embryogenesis and carcinogenesis. Acta Histochem 2016; 118: 693-703.
  14. Drew DA, Cao Y, Chan AT: Aspirin and colorectal cancer: the promise of precision chemoprevention. Nat Rev Cancer 2016; 16: 173-186.
  15. Usman MW, Luo F, Cheng H, Zhao JJ, Liu P: Chemopreventive effects of aspirin at a glance. Biochim Biophys Acta 2015; 1855: 254-263.
  16. Tian Y, Ye Y, Gao W, Chen H, Song T, Wang D, Mao X, Ren C: Aspirin promotes apoptosis in a murine model of colorectal cancer by mechanisms involving downregulation of IL-6-STAT3 signaling pathway. Int J Colorectal Dis 2011; 26: 13-22.
  17. Ding JH, Yuan LY, Huang RB, Chen GA: Aspirin inhibits proliferation and induces apoptosis of multiple myeloma cells through regulation of Bcl-2 and Bax and suppression of VEGF. Eur J Haematol 2014; 93: 329-339.
  18. Liao D, Zhong L, Duan T, Zhang RH, Wang X, Wang G, Hu K, Lv X, Kang T: Aspirin Suppresses the Growth and Metastasis of Osteosarcoma through the NF-kappaB Pathway. Clin Cancer Res 2015; 21: 5349-5359.
  19. Rothwell PM, Wilson M, Price JF, Belch JF, Meade TW, Mehta Z: Effect of daily aspirin on risk of cancer metastasis: a study of incident cancers during randomised controlled trials. Lancet 2012; 379: 1591-1601.
  20. Ogawa F, Amano H, Ito Y, Matsui Y, Hosono K, Kitasato H, Satoh Y, Majima M: Aspirin reduces lung cancer metastasis to regional lymph nodes. Biomed Pharmacother 2014; 68: 79-86.
  21. Shi C, Zhang N, Feng Y, Cao J, Chen X, Liu B: Aspirin Inhibits IKK-beta-mediated Prostate Cancer Cell Invasion by Targeting Matrix Metalloproteinase-9 and Urokinase-Type Plasminogen Activator. Cell Physiol Biochem 2017; 41: 1313-1324.
  22. Moon CM, Kwon JH, Kim JS, Oh SH, Jin Lee K, Park JJ, Pil Hong S, Cheon JH, Kim TI, Kim WH: Nonsteroidal anti-inflammatory drugs suppress cancer stem cells via inhibiting PTGS2 (cyclooxygenase 2) and NOTCH/ HES1 and activating PPARG in colorectal cancer. Int J Cancer 2014; 134: 519-529.
  23. Bratasz A, Selvendiran K, Wasowicz T, Bobko A, Khramtsov VV, Ignarro LJ, Kuppusamy P: NCX-4040, a nitric oxide-releasing aspirin, sensitizes drug-resistant human ovarian xenograft tumors to cisplatin by depletion of cellular thiols. J Transl Med 2008; 6: 9.
  24. Kopp E, Ghosh S: Inhibition of NF-kappa B by sodium salicylate and aspirin. Science 1994; 265: 956-959.
  25. McCarty MF, Block KI: Preadministration of high-dose salicylates, suppressors of NF-kappaB activation, may increase the chemosensitivity of many cancers: an example of proapoptotic signal modulation therapy. Integr Cancer Ther 2006; 5: 252-268.
  26. Pan MR, Chang HC, Hung WC: Non-steroidal anti-inflammatory drugs suppress the ERK signaling pathway via block of Ras/c-Raf interaction and activation of MAP kinase phosphatases. Cell Signal 2008; 20: 1134-1141.
  27. Bos CL, Kodach LL, van den Brink GR, Diks SH, van Santen MM, Richel DJ, Peppelenbosch MP, Hardwick JC: Effect of aspirin on the Wnt/beta-catenin pathway is mediated via protein phosphatase 2A. Oncogene 2006; 25: 6447-6456.
  28. Guo ZK, Guo K, Luo H, Mu LM, Li Q, Chang YQ: The expression analysis of nanog in the developing rat myocardial tissues. Cell Physiol Biochem 2015; 35: 866-874.
  29. Nagata T, Shimada Y, Sekine S, Hori R, Matsui K, Okumura T, Sawada S, Fukuoka J, Tsukada K: Prognostic significance of NANOG and KLF4 for breast cancer. Breast Cancer 2014; 21: 96-101.
  30. Lee M, Nam EJ, Kim SW, Kim S, Kim JH, Kim YT: Prognostic impact of the cancer stem cell-related marker NANOG in ovarian serous carcinoma. Int J Gynecol Cancer 2012; 22: 1489-1496.
  31. Meng HM, Zheng P, Wang XY, Liu C, Sui HM, Wu SJ, Zhou J, Ding YQ, Li J: Over-expression of Nanog predicts tumor progression and poor prognosis in colorectal cancer. Cancer Biol Ther 2010; 9: 295-302.
  32. Zhang J, Espinoza LA, Kinders RJ, Lawrence SM, Pfister TD, Zhou M, Veenstra TD, Thorgeirsson SS, Jessup JM: NANOG modulates stemness in human colorectal cancer. Oncogene 2013; 32: 4397-4405.
  33. Ibrahim EE, Babaei-Jadidi R, Saadeddin A, Spencer-Dene B, Hossaini S, Abuzinadah M, Li N, Fadhil W, Ilyas M, Bonnet D, Nateri AS: Embryonic NANOG activity defines colorectal cancer stem cells and modulates through AP1- and TCF-dependent mechanisms. Stem Cells 2012; 30: 2076-2087.
  34. Han J, Zhang F, Yu M, Zhao P, Ji W, Zhang H, Wu B, Wang Y, Niu R: RNA interference-mediated silencing of NANOG reduces cell proliferation and induces G0/G1 cell cycle arrest in breast cancer cells. Cancer Lett 2012; 321: 80-88.
  35. Chiou SH, Wang ML, Chou YT, Chen CJ, Hong CF, Hsieh WJ, Chang HT, Chen YS, Lin TW, Hsu HS, Wu CW: Coexpression of Oct4 and Nanog enhances malignancy in lung adenocarcinoma by inducing cancer stem cell-like properties and epithelial-mesenchymal transdifferentiation. Cancer Res 2010; 70: 10433-10444.
  36. Siu MKY, Wong ESY, Kong DSH, Chan HY, Jiang L, Wong OGW, Lam EWF, Chan KKL, Ngan HYS, Le XF, Cheung AN: Stem cell transcription factor NANOG controls cell migration and invasion via dysregulation of E-cadherin and FoxJ1 and contributes to adverse clinical outcome in ovarian cancers. Oncogene 2013; 32: 3500-3509.
  37. Hu H, Zhang S, Zhu S: Influence of aspirin and cigarette smoke extract on the expression of cyclin D1 and effects of cell cycle in esophageal squamous cell carcinoma cell line. Dis Esophagus 2009; 22: 310-316.
  38. Khan P, Manna A, Saha S, Mohanty S, Mukherjee S, Mazumdar M, Guha D, Das T: Aspirin inhibits epithelial-to-mesenchymal transition and migration of oncogenic K-ras-expressing non-small cell lung carcinoma cells by down-regulating E-cadherin repressor Slug. BMC Cancer 2016; 16: 39.
  39. Dovizio M, Bruno A, Tacconelli S, Patrignani P: Mode of action of aspirin as a chemopreventive agent. Recent Results Cancer Res 2013; 191: 39-65.
  40. Douville J, Beaulieu R, Balicki D: ALDH1 as a functional marker of cancer stem and progenitor cells. Stem Cells Dev 2009; 18: 17-25.
  41. Gabay M, Li Y, Felsher DW: MYC activation is a hallmark of cancer initiation and maintenance. Cold Spring Harb Perspect Med 2014; 4.
  42. Zeineddine D, Hammoud AA, Mortada M, Boeuf H: The Oct4 protein: more than a magic stemness marker. Am J Stem Cells 2014; 3: 74-82.
    External Resources
  43. Boumahdi S, Driessens G, Lapouge G, Rorive S, Nassar D, Le Mercier M, Delatte B, Caauwe A, Lenglez S, Nkusi E, Brohee S, Salmon I, Dubois C, del Marmol V, Fuks F, Beck B, Blanpain C: SOX2 controls tumour initiation and cancer stem-cell functions in squamous-cell carcinoma. Nature 2014; 511: 246-250.
  44. Yin M, Yuan Y, Cui Y, Hong X, Luo H, Hu X, Tang M, Hescheler J, Xi J: Puerarin Suppresses the Self-Renewal of Murine Embryonic Stem Cells by Inhibition of REST-MiR-21 Regulatory Pathway. Cell Physiol Biochem 2015; 37: 527-536.
  45. Chen B, Dodge ME, Tang W, Lu J, Ma Z, Fan CW, Wei S, Hao W, Kilgore J, Williams NS, Roth MG, Amatruda JF, Chen C, Lum L: Small molecule-mediated disruption of Wnt-dependent signaling in tissue regeneration and cancer. Nat Chem Biol 2009; 5: 100-107.
  46. Thorne CA, Hanson AJ, Schneider J, Tahinci E, Orton D, Cselenyi CS, Jernigan KK, Meyers KC, Hang BI, Waterson AG, Kim K, Melancon B, Ghidu VP, Sulikowski GA, LaFleur B, Salic A, Lee LA, Miller DM, 3rd, Lee E: Small-molecule inhibition of Wnt signaling through activation of casein kinase 1alpha. Nat Chem Biol 2010; 6: 829-836.
  47. Li Y, Wicha MS, Schwartz SJ, Sun D: Implications of cancer stem cell theory for cancer chemoprevention by natural dietary compounds. J Nutr Biochem 2011; 22: 799-806.
  48. Kakarala M, Brenner DE, Korkaya H, Cheng C, Tazi K, Ginestier C, Liu S, Dontu G, Wicha MS: Targeting breast stem cells with the cancer preventive compounds curcumin and piperine. Breast Cancer Res Treat 2010; 122: 777-785.
  49. Li Y, Zhang T, Korkaya H, Liu S, Lee HF, Newman B, Yu Y, Clouthier SG, Schwartz SJ, Wicha MS, Sun D: Sulforaphane, a dietary component of broccoli/broccoli sprouts, inhibits breast cancer stem cells. Clin Cancer Res 2010; 16: 2580-2590.
  50. Hirsch HA, Iliopoulos D, Struhl K: Metformin inhibits the inflammatory response associated with cellular transformation and cancer stem cell growth. Proc Natl Acad Sci U S A 2013; 110: 972-977.
  51. Smith T, Elwood P, Keating C, Rothwell P, Detering E, Freedman A, Langley R, Logan R, Phillips C, DeCensi A: The Aspirin Foundation Scientific Conference: the history, the present state and the future of aspirin prophylaxis. Ecancermedicalscience 2014; 8: 388.
  52. Ricchi P, Pignata S, Di Popolo A, Memoli A, Apicella A, Zarrilli R, Acquaviva AM: Effect of aspirin on cell proliferation and differentiation of colon adenocarcinoma Caco-2 cells. Int J Cancer 1997; 73: 880-884.
  53. Supino R, Mazzoni A, Formelli F: Effect of medroxyprogesterone acetate and of some antiinflammatory agents on mouse erythroleukemia cell differentiation. Tumori 1984; 70: 17-21.
    External Resources

Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: May 28, 2017
Accepted: September 22, 2017
Published online: November 27, 2017
Issue release date: December 2017

Number of Print Pages: 13
Number of Figures: 6
Number of Tables: 0

ISSN: 1015-8987 (Print)
eISSN: 1421-9778 (Online)

For additional information: https://www.karger.com/CPB

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