Biotechnology Letters

, Volume 28, Issue 16, pp 1263–1270 | Cite as

Cyclooxygenase-independent effects of aspirin on HT-29 human colon cancer cells, revealed by oligonucleotide microarrays

  • Hongying Yin
  • Hao Xu
  • Yongchao Zhao
  • Weiping Yang
  • Jing Cheng
  • Yuxiang ZhouEmail author
Original Paper


Aspirin and other non-steroidal anti-inflammatory drugs (NSAIDs) inhibit proliferation of human colon cancer cells in vitro. Transmission electron microscope detected morphological features of apoptosis in the aspirin-treated (5 mM, 72 h) HT-29 cells in which cyclooxygenoase-2 is catalytically inactive. We investigated aspirin-induced genome-wide expression changes in HT-29 cells and further studied the time- and concentration-dependent expression changes in 374 apoptosis-related genes, which is the first to show stimulation of genome-wide expression of HT-29 cells by aspirin. The most marked effects of aspirin are on ribosome assembly and rRNA metabolism, which could explain why the quasi-apoptotic morphological changes are not accompanied by a classical DNA ladder. These findings demonstrate that aspirin induces apoptosis in HT-29 cells, bolstering the hypothesis that apoptosis may be a mechanism by which NSAIDs inhibit colon carcinogenesis.


Aspirin Apoptosis Colon cancer Cyclooxygenase-2 (COX-2) DNA microarray 


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We thank Ch. Zhou, W. Shao, and L. Wang for helpful discussions and comments on the manuscript; Dr. D. Wright for his assistance with manuscript presentation; Dr. H. Smith for her review of the manuscript. This work was supported by grants from the Beijing Natural Science Foundation of China (No. H010210640121) and State Key Laboratory of Natural and Biomimetic Drugs (No. 0200006209089112565).


  1. Amin AR, Vyas P, Attur M, Leszczynska-Piziak J, Patel IR, Weissmann G, Abramson SB (1995) The mode of action of aspirin-like drugs: effect on inducible nitric oxide synthase. Proc Natl Acad Sci USA 92:7926–7930PubMedCrossRefGoogle Scholar
  2. Baron JA, Cole BF, Sandler RS, Haile RW, Ahnen D, Bresalier R, McKeown-Eyssen G, Summers RW, Rothstein R, Burke CA, Snover DC, Church TR, Allen JI, Beach M, Beck GJ, Bond JH, Byers T, Greenberg ER, Mandel JS, Marcon N, Mott LA, Pearson L, Saibil F, van Stolk RU (2003) A randomized trial of aspirin to prevent colorectal adenomas. N Engl J Med 348:891–899PubMedCrossRefGoogle Scholar
  3. Castano E, Dalmau M, Barragan M, Pueyo G, Bartrons R, Gil J (1999) Aspirin induces cell death and caspase-dependent phosphatidylserine externalization in HT-29 human colon adenocarcinoma cells. Br J Cancer 81:294–299PubMedCrossRefGoogle Scholar
  4. Chan AT, Giovannucci EL, Schernhammer ES, Colditz GA, Hunter DJ, Willett WC, Fuchs CS (2004) A prospective study of aspirin use and the risk for colorectal adenoma. Ann Intern Med 140:157–166PubMedGoogle Scholar
  5. Dahlquist KD, Salomonis N, Vranizan K, Lawlor SC, Conklin BR (2002) GenMAPP, a new tool for viewing and analyzing microarray data on biological pathways. Nat Genet 31:19–20PubMedCrossRefGoogle Scholar
  6. Fenwick SW, Toogood GJ, Lodge JP, Hull MA (2003) The effect of the selective cyclooxygenase-2 inhibitor rofecoxib on human colorectal cancer liver metastases. Gastroenterology 125:716–729PubMedCrossRefGoogle Scholar
  7. Gagna CE, Winokur D, Clark Lambert W (2004) Cell biology, chemogenomics and chemoproteomics. Cell Biol Int 28:755–764PubMedCrossRefGoogle Scholar
  8. Giovannucci E, Rimm EB, Stampfer MJ, Colditz GA, Ascherio A, Willett WC (1994) Aspirin use and the risk for colorectal cancer and adenoma in male health professionals. Ann Intern Med 121:241–246PubMedGoogle Scholar
  9. Giovannucci E, Egan KM, Hunter DJ, Stampfer MJ, Coldtiz GA, Willett WC, Speizwer FE (1995) Aspirin and the risk of colorectal cancer in woman. N Engl J Med 333:609–614PubMedCrossRefGoogle Scholar
  10. Hardwick JC, Van Santen M, Van Den Brink GR, Van Deventer SJ, Peppelenbosch MP (2004) DNA array analysis of the effects of aspirin on colon cancer cells: involvement of Rac1. Carcinogenesis 25:1293–1298PubMedCrossRefGoogle Scholar
  11. Henry D, Dobson A, Turner C, Hall P, Forbes C, Patey P (1991) NSAIDs and risk of upper gastrointestinal bleeding. Lancet 337:730PubMedCrossRefGoogle Scholar
  12. Hsi LC, Baek SJ, Eling TE (2000) Lack of cyclooxygenase-2 activity in HT-29 human colorectal carcinoma cells. Exp Cell Res 256:563–570PubMedCrossRefGoogle Scholar
  13. Lynch S, Brogden RN (1986) Etodolac. A preliminary review of its pharmacodynamic activity and therapeutic use. Drugs 31:288–300PubMedCrossRefGoogle Scholar
  14. Marton MJ, DeRisi JL, Bennett HA, Iyer VR, Meyer MR, Roberts CJ, Stoughton R, Burchard J, Slade D, Dai H, Bassett Jr DE, Hartwell LH, Brown PO, Friend SH (1998) Drug target validation and identification of secondary drug target effects using DNA microarrays. Nat Med 4:1293–1301PubMedCrossRefGoogle Scholar
  15. Marx J (2001) Cancer research. Anti-inflammatories inhibit cancer growth – but how? Science 291:581–582PubMedCrossRefGoogle Scholar
  16. Qiao L, Hanif R, Sphicas E, Shiff SJ, Rigas B (1998) Effect of aspirin on induction of apoptosis in HT-29 human colon adenocarcinoma cells. Biochem Pharmacol 55:53–64PubMedCrossRefGoogle Scholar
  17. Rahme E, Barkun AN, Toubouti Y, Bardou M (2003) The cyclooxygenase-2-selective inhibitors rofecoxib and celecoxib prevent colorectal neoplasia occurrence and recurrence. Gastroenterology 125:404–412PubMedCrossRefGoogle Scholar
  18. Reddy BS, Rao CV, Rivenson A, Kelloff G (1993) Inhibitory effect of aspirin on azoxymethane-induced colon carcinogenesis in F344 rats. Carcinogenesis 14:1493–1497PubMedCrossRefGoogle Scholar
  19. Ricchi P, Pignata S, Di Popolo A, Memoli A, Apicella A, Zarrilli R, Acquaviva AM (1997) Effect of aspirin on cell proliferation and differentiation of colon adenocarcinoma Caco-2 cells. Int J Cancer 73:880–884PubMedCrossRefGoogle Scholar
  20. Rigas B, Goldman IS, Levine L (1993) Altered eicosanoid levels in human colon cancer. J Lab Clin Med 122:518–523PubMedGoogle Scholar
  21. Ruschoff J, Wallinger S, Dietmaier W, Bocker T, Brockhoff G, Hofstadter F, Fishel R (1998) Aspirin suppresses the mutator phenotype associated with hereditary nonpolyposis colorectal cancer by genetic selection. Proc Natl Acad Sci USA 95:11301–11306PubMedCrossRefGoogle Scholar
  22. Sakon S, Xue X, Takekawa M, Sasazuki T, Okazaki T, Kojima Y, Piao JH, Yagita H, Okumura K, Doi T, Nakano H (2003) NF-kappaB inhibits TNF-induced accumulation of ROS that mediate prolonged MAPK activation and necrotic cell death. EMBO J 22:3898–3909PubMedCrossRefGoogle Scholar
  23. Schernhammer ES, Kang JH, Chan AT, Michaud DS, Skinner HG, Giovannucci E, Colditz GA, Fuchs CS (2004) A prospective study of aspirin use and the risk of pancreatic cancer in women. J Natl Cancer Inst 96:22–28PubMedGoogle Scholar
  24. Smith ML, Hawcroft G, Hull MA (2000) The effect of non-steroidal anti-inflammatory drugs on human colorectal cancer cells: evidence of different mechanisms of action. Eur J Cancer 36:664–674PubMedCrossRefGoogle Scholar
  25. Stark LA, Din FV, Zwacka RM, Dunlop MG (2001) Aspirin-induced activation of the NF-kappaB signaling pathway: a novel mechanism for aspirin-mediated apoptosis in colon cancer cells. FASEB J 15:1273–1275PubMedGoogle Scholar
  26. Subbegowda R, Frommel T (1998) Aspirin toxicity for human colonic tumor cells results from necrosis and is accompanied by cell cycle arrest. Cancer Res 58:2772–2776PubMedGoogle Scholar
  27. Thun MJ, Namboodiri MM, Calle EE, Flanders WD, Heath Jr CW (1993) Aspirin use and risk of fatal cancer. Cancer Res 53:1322–1327PubMedGoogle Scholar
  28. Totzke G, Schulze-Osthoff K, Janicke RU (2003) Cyclooxygenase-2 (COX-2) inhibitors sensitize tumor cells specifically to death receptor-induced apoptosis independently of COX-2 inhibition. Oncogene 22:8021–8030PubMedCrossRefGoogle Scholar
  29. Van Gelder RN, von Zastrow ME, Yool A, Dement WC, Barchas JD, Eberwine JH (1990) Amplified RNA synthesized from limited quantities of heterogeneous cDNA. Proc Natl Acad Sci USA 87:1663–1667PubMedCrossRefGoogle Scholar
  30. Vanchieri C (2004) Vioxx withdrawal alarms cancer prevention researchers. J Natl Cancer Inst 96:1734–1735PubMedCrossRefGoogle Scholar
  31. Wang D, Mann JR, DuBois RN (2005) The role of prostaglandins and other eicosanoids in the gastrointestinal tract. Gastroenterology 128:1445–1461PubMedCrossRefGoogle Scholar
  32. Watson AJ (2006) An overview of apoptosis and the prevention of colorectal cancer. Crit Rev Oncol Hematol 57:107–121PubMedCrossRefGoogle Scholar
  33. Yamomoto Y, Yin MJ, Lin KM, Gaynor RB (1999) Sulindac inhibits activation of the NF-kappaB pathway. J Biol Chem 274:27307–27314CrossRefGoogle Scholar
  34. Yang YY, Hu CJ, Chang SM, Tai TY, Leu SJ (2004) Aspirin inhibits monocyte chemoattractant protein-1 and interleukin-8 expression in TNF-alpha stimulated human umbilical vein endothelial cells. Atherosclerosis 174:207–213PubMedGoogle Scholar
  35. Yin MJ, Yamamoto Y, Gaynor RB (1998) The anti-inflammatory agents aspirin and salicylate inhibit the activity of I(kappa)B kinase-beta. Nature 396:77–80PubMedCrossRefGoogle Scholar
  36. Zhang L, Yu J, Park BH, Kinzler KW, Vogelstein B (2000) Role of BAX in the apoptotic response to anticancer agents. Science 290:989–992PubMedCrossRefGoogle Scholar
  37. Zhang L, Zhang Y, Zhou Y, An S, Cheng J (2002) Response of gene expression in Saccharomyces cerevisiae to amphotericin B and nystatin measured by microarrays. J Antimicrob Chemother 49:905–915PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Hongying Yin
    • 1
    • 2
    • 3
  • Hao Xu
    • 4
  • Yongchao Zhao
    • 1
    • 2
    • 3
  • Weiping Yang
    • 4
  • Jing Cheng
    • 1
    • 2
    • 3
  • Yuxiang Zhou
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of Biological Sciences and BiotechnologyTsinghua UniversityBeijingChina
  2. 2.Medical Systems Biology Research CenterTsinghua University, School of MedicineBeijingChina
  3. 3.National Engineering Research Center for Beijing Biochip Technology (NERCBBT)BeijingChina
  4. 4.CapitalBio CorporationBeijingChina

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