, Volume 14, Issue 7, pp 913–922 | Cite as

Sulindac induces apoptotic cell death in susceptible human breast cancer cells through, at least in part, inhibition of IKKβ

  • A-Mi Seo
  • Seung-Woo Hong
  • Jae-Sik Shin
  • In-Chul Park
  • Nam-Joo Hong
  • Dae-Jin Kim
  • Won-Keun Lee
  • Wang-Jae Lee
  • Dong-Hoon JinEmail author
  • Myeong-Sok LeeEmail author
Original Paper


Sulindac is a non-steroidal anti-inflammatory agent with anti-tumor activities that include the induction of apoptosis in various cancer cells and the inhibition malignant transformation. However, the molecular mechanisms underlying these effects are unclear. Recently, it has been shown that sulindac can inhibit NF-κB activation. Here, we demonstrate that sulindac induces apoptotic cell death in susceptible human breast cancer cells through, at least in part, inhibition of IKKβ activity. More specifically, when we compared two different human breast cancer cell lines, Hs578T, which has relatively low basal IKKβ activity, and MDA-MB231, which has relatively high basal IKKβ activity, we found that MDA-MB231 was markedly more sensitive to sulindac-induced apoptosis than Hs578T. This was associated with greater caspase-3 and -9 activity in sulindac-treated MDA-MB231 cells. Using a combination of chemical kinase inhibitors and siRNA-mediated knockdown of specific kinases, we found that sulindac inhibits IKKβ, which, in turn, leads to the p38 MAPK-dependent activation of JNK1. Together, these findings suggest that sulindac induces apoptosis in susceptible human breast cancer cells through, at least in part, the inhibition of IKKβ and the subsequent p38 MAPK-dependent activation of JNK1.


NSAIDs Sulindac IKKβ JNK1 P38/MAPK Apoptosis 



Non-steroidal anti-inflammatory drugs


IκB kinase β


c-Jun NH2-terminal kinase 1




Matogen-activated protein kinase


Nuclear factor kappa B


Inhibitory kappa B



Financial support: This research was supported by the Research Center for Women’s Diseases, at the Korea Science and Engineering Foundation (KOSEF) and by a grant from Sookmyung Women’s University.

Supplementary material

10495_2009_367_MOESM1_ESM.pdf (61 kb)
Supplementary material 1 (PDF 60 kb)


  1. 1.
    Piazza GA, Rahm AL, Krutzsch M, Sperl G, Paranka NS, Gross PH et al (1995) Antineoplastic drugs sulindac sulfide and sulfone inhibit cell growth by inducing apoptosis. Cancer Res 55:3110–3116PubMedGoogle Scholar
  2. 2.
    Lim JT, Piazza GA, Han EK, Delohery TM, Li H, Finn TS et al (1999) Sulindac derivatives inhibit growth and induce apoptosis in human prostate cancer cell line. Biochem Pharmacol 58(7):1097–1107. doi: 10.1016/S0006-2952(99)00200-2 PubMedCrossRefGoogle Scholar
  3. 3.
    Ikui AE, Yao Y, Zhou P, Weinstein IB (2001) Induction of apoptosis by sulindac sulfide in HL60 cells is enhanced by p21CiP1 or p27KiP1. Anticancer Res 21:2297–2303PubMedGoogle Scholar
  4. 4.
    Adachi M, Sakamoto H, Kawamura R, Wang W, Imai K, Shinomura Y (2007) Nonsteroidal anti-inflammatory drugs and oxidative stress in cancer cells. Histol Histopathol 22:437–442PubMedGoogle Scholar
  5. 5.
    Shiff SJ, Qiao L, Tsai LL, Rigas B (1995) Sulindac sulfide, an aspirin-like compound, inhibits proliferation, causes cell cycle quiescence, and induces apoptosis in HT-29 colon adenocarcinoma cells. J Clin Invest 96:491–503. doi: 10.1172/JCI.118060 PubMedCrossRefGoogle Scholar
  6. 6.
    Lee HC, Park IC, Park MJ, An S, Woo SH, Jin HO et al (2005) Sulindac and its metabolites inhibit invasion of glioblastoma cells via down-regulation of Akt/PKB and MMP-2. J Cell Biochem 94(3):597–610. doi: 10.1002/jcb.20312 PubMedCrossRefGoogle Scholar
  7. 7.
    Reddy BS, Rao CV, Seibert K (1996) Evaluation of cyclooxygenase-2 inhibitor for potential chemopreventive properties in colon carcinogenesis. Cancer Res 56:4566–4569PubMedGoogle Scholar
  8. 8.
    Narayanan BA, Narayanan NK, Pittman B, Reddy BS (2004) Regression of mouse prostatic intraepithelial neoplasia by nonsteroidal anti-inflammatory drugs in the transgenic adenocarcinoma mouse prostate model. Clin Cancer Res 10:7727–7737. doi: 10.1158/1078-0432.CCR-04-0732 PubMedCrossRefGoogle Scholar
  9. 9.
    Yamamato Y, Yin M, Ling K, Gaynor RB (1999) Sulindac inhibits activation of the NF-κB pathway. J Biol Chem 274:27307–27314. doi: 10.1074/jbc.274.38.27307 CrossRefGoogle Scholar
  10. 10.
    Wu MX, Ao Z, Prasad KV, Wu R, Schlossman SX (1998) IEX-1L, an apoptosis inhibitor involved in NF-κB-mediated cell survival. Science 281:998–1001. doi: 10.1126/science.281.5379.998 PubMedCrossRefGoogle Scholar
  11. 11.
    Baldwin AS (2001) Control of oncogenesis and cancer therapy resistance by the transcription factor NF-κB. J Clin Invest 107:241–246. doi: 10.1172/JCI.11991 PubMedCrossRefGoogle Scholar
  12. 12.
    Kwak YT, Guo J, Shen J, Gaynor RB (2000) Analysis of domains in the IKKalpha and IKKbeta proteins that regulate their kinase activity. J Biol Chem 275:14752–14759. doi: 10.1074/jbc.M001039200 PubMedCrossRefGoogle Scholar
  13. 13.
    Mercurio F, Zhu H, Murray BW, Shevchenko A, Bennett BL, Li J et al (1997) IKK-1 and IKK-2: cytokine-activated IkappaB kinases essential for NF-kappaB activation. Science 278(5339):860–866. doi: 10.1126/science.278.5339.860 PubMedCrossRefGoogle Scholar
  14. 14.
    Didonato JA, Hayakawa M, Rothwarf DM, Zandi E, Karin M (1997) A cytokine-responsive IkappaB kinase that activates the transcription factor NF-kappaB. Nature 388:548–554. doi: 10.1038/41493 PubMedCrossRefGoogle Scholar
  15. 15.
    Beg AA, Ruben SM, Scheinman RI, Haskill S, Rosen CA, Baldwin AS (1992) I kappa B interacts with the nuclear localization sequences of the subunits of NF-kappa B: a mechanism for cytoplasmic retention. Genes Dev 6:1899–1913. doi: 10.1101/gad.6.10.1899 PubMedCrossRefGoogle Scholar
  16. 16.
    Brown K, Gerstberger S, Carlson L, Fransozo G, Siebenlist U (1995) Control of I kappa B-alpha proteolysis by site-specific, signal-induced phosphorylation. Science 267:1485–1488. doi: 10.1126/science.7878466 PubMedCrossRefGoogle Scholar
  17. 17.
    Li Q, Lu Q, Hwang JY, Buscher D, Lee KF, Izpisua-Belmonte JC, Verma IM (1999) IKK1-deficient mice exhibit abnormal development of skin and skeleton. Genes Dev 13:1322–1328. doi: 10.1101/gad.13.10.1322 PubMedCrossRefGoogle Scholar
  18. 18.
    Li ZW, Chu W, Hu Y, Delhase M, Deerinck T, Ellisman R, Karin M (1999) The IKKbeta subunit of IkappaB kinase (IKK) is essential for nuclear factor kappaB activation and prevention of apoptosis. J Exp Med 189:1839–1845. doi: 10.1084/jem.189.11.1839 PubMedCrossRefGoogle Scholar
  19. 19.
    Tanaka H, Fujita N, Tsuruo T (2005) 3-Phosphoinositide-dependent protein kinase-1-mediated IkappaB kinase beta (IkkB) phosphorylation activates NF-kappaB signaling. J Biol Chem 280:40965–40973. doi: 10.1074/jbc.M506235200 PubMedCrossRefGoogle Scholar
  20. 20.
    Davis RJ (2000) Signal transduction by the JNK group of MAP kinases. Cell 103(2):239–252. doi: 10.1016/S0092-8674(00)00116-1 PubMedCrossRefGoogle Scholar
  21. 21.
    Chang L, Karin M (2001) Mammalian MAP kinas signaling cascades. Nature 410(6824):37–40. doi: 10.1038/35065000 PubMedCrossRefGoogle Scholar
  22. 22.
    Kim TI, Jin SH, Kim WH, Kang EH, Choi KY, Kim HJ et al (2001) Prolonged activation of mitogen-activated protein kinases during NSAID-induced apoptosis in HT-29 colon cancer cells. Int J Colorectal Dis 16:167–173. doi: 10.1007/s003840100301 PubMedCrossRefGoogle Scholar
  23. 23.
    Lim SJ, Lee YJ, Park DH, Lee E, Choi MK, Park W et al (2007) Alpha-tocopheryl succinate sensitizes human colon cancer cells to exisulind-induced apoptosis. Apoptosis 12:423–431. doi: 10.1007/s10495-006-0620-9 PubMedCrossRefGoogle Scholar
  24. 24.
    Song Z, Tong C, Liang J, Dockendorff A, Huang C, Augenlicht LH, Yang W (2007) JNK1 is required for sulindac-mediated inhibition of cell proliferation and induction of apoptosis in vitro and in vivo. Eur J Pharm 560:95–100. doi: 10.1016/j.ejphar.2007.01.020 CrossRefGoogle Scholar
  25. 25.
    Dong C, Yang DD, Wysk M, Whimarsh AJ, Davis RJ, Flavell RA (1998) Defective T cell differentiation in the absence of JNK1. Science 282:2092–2095. doi: 10.1126/science.282.5396.2092 PubMedCrossRefGoogle Scholar
  26. 26.
    Liu J, Minemoto Y, Lin A (2004) c-Jun N-terminal protein kinase 1 (JNK1), but not JNK2, is essential for tumor necrosis factor alpha-induced c-Jun kinase activation and apoptosis. Mol Cell Biol 24:10844–10856. doi: 10.1128/MCB.24.24.10844-10856.2004 PubMedCrossRefGoogle Scholar
  27. 27.
    Seo SK, Lee HC, Woo SH, Jin HO, Yoo DH, Lee SJ et al (2007) Sulindac-derived reactive oxygen species induce apoptosis of human multiple myeloma cells via p38 mitogen activated protein kinase-induced mitochondrial dysfunction. Apoptosis 12:195–209. doi: 10.1007/s10495-006-0527-5 PubMedCrossRefGoogle Scholar
  28. 28.
    Minami T, Adachi M, Kawamura R, Zhang Y, Shinomura Y, Imai K (2005) Sulindac enhances the proteasome inhibitor bortezomib-mediated oxidative stress and anticancer activity. Clin Cancer Res 11:5248–5256. doi: 10.1158/1078-0432.CCR-05-0085 PubMedCrossRefGoogle Scholar
  29. 29.
    Tegeder I, Pfeilschifter J, Geisslinger G (2001) Cyclooxygenase-independent actions of cyclooxygenase inhibitors. FASEB J 15:2057–2072. doi: 10.1096/fj.01-0390rev PubMedCrossRefGoogle Scholar
  30. 30.
    Sun Y, Sinicrope FA (2005) Selective inhibitors of MEK1/ERK44/42 and p38 mitogen-activated protein kinases potentiate apoptosis induction by sulindac sulfide in human colon carcinoma cells. Mol Cancer Ther 4:51–59PubMedGoogle Scholar
  31. 31.
    Alpert D, Schwenger P, Han J, Vilcek J (1999) Cell stress and MKK6b-mediated p38 MAP kinase activation inhibit tumor necrosis factor-induced IkB phosphorylation and NF-kB activation. J Biol Chem 274:22176–22183. doi: 10.1074/jbc.274.32.22176 PubMedCrossRefGoogle Scholar
  32. 32.
    Ivanov VN, Ronai Z (2000) p38 protects human melanoma cells from UV-induced apoptosis through down-regulation of NF-kB activity and Fas expression. Oncogene 19:3003–3012. doi: 10.1038/sj.onc.1203602 PubMedCrossRefGoogle Scholar
  33. 33.
    Monks NR, Pardee AB (2006) Targeting the NF-kB pathway in estrogen receptor negative MDA-MB-231 breast cancer cells using small inhibitory RNAs. J Cell Biochem 98:221–233. doi: 10.1002/jcb.20789 PubMedCrossRefGoogle Scholar
  34. 34.
    Oleinik NV, Krupenko NI, Krupenko SA (2007) Cooperation between JNK1 and JNK2 in activation of p53 apoptotic pathway. Oncogene 26:7222–7230. doi: 10.1038/sj.onc.1210526 PubMedCrossRefGoogle Scholar
  35. 35.
    Han EK, Arber N, Yamamoto H, Lim JT, Delohery T, Pamukcu R et al (1998) Effects of sulindac and its metabolites on growth and apoptosis in human mammary epithelial and breast carcinoma cell lines. Breast Cancer Res Treat 48:195–203. doi: 10.1023/A:1005924730450 PubMedCrossRefGoogle Scholar
  36. 36.
    Lee DF, Kuo HP, Chen CT, Hsu JM, Chou CK, Wei YK et al (2007) IKKβ suppression of TSC1 links inflammation and tumor angiogenesis via the mTOR pathway. Cell 130(3):440–455. doi: 10.1016/j.cell.2007.05.058 PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • A-Mi Seo
    • 1
  • Seung-Woo Hong
    • 2
    • 3
    • 4
  • Jae-Sik Shin
    • 3
    • 4
  • In-Chul Park
    • 5
  • Nam-Joo Hong
    • 6
  • Dae-Jin Kim
    • 7
  • Won-Keun Lee
    • 2
  • Wang-Jae Lee
    • 3
    • 4
  • Dong-Hoon Jin
    • 3
    Email author
  • Myeong-Sok Lee
    • 1
    Email author
  1. 1.Research Center for Women’s Diseases, Division of Biological SciencesSookmyung Women’s UniversitySeoulKorea
  2. 2.Department of Biological SciencesMyongji UniversityYonginKorea
  3. 3.Department of Anatomy and Tumor Immunity Medical Research CenterSeoul National University College of MedicineSeoulKorea
  4. 4.Cancer Research InstituteSeoul National University College of MedicineSeoulKorea
  5. 5.Laboratory of Functional GenomicsKorea Institute of Radiological and Medical SciencesSeoulKorea
  6. 6.School of BiotechnologyYeungnam UniversityDaedong, Gyungsan CityKorea
  7. 7.Department of Anatomy, College of MedicineChung-Ang UniversitySeoulRepublic of Korea

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