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Celecoxib induces apoptosis in cervical cancer cells independent of cyclooxygenase using NF-κB as a possible target

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Abstract

Purpose

Recently, many studies have shown that celecoxib induces apoptosis in various cancer cells by different mechanisms depending on the cell type. This study examined the apoptotic effect of celecoxib in cervical cancer cells and its mechanism.

Methods

Cell viability was measured by MTT assay and apoptosis was examined by DNA fragmentation and flow cytometry. Western blotting and immunoprecipitation were used to explore various mechanisms of celecoxib-induced apoptosis. The activation of NF-κB was confirmed by EMSA.

Results

Celecoxib induced apoptosis independent of COX-2 activity. This event accompanied the activation of caspase-8 and -9 with Bid cleavage and the loss of mitochondrial membrane potential. The protective effect of caspase-8 and -9 inhibitors on celecoxib-induced apoptosis suggests the importance of caspase-8 and -9 activation in this apoptotic pathway. Fas/FADD-mediated apoptotic pathway was detected only in C33A cells, demonstrated by the immunoprecipitation of Fas-FADD in celecoxib-treated cells and the protective effect of FADD dominant negative mutant. Finally, NF-κB appeared to be involved in celecoxib-induced apoptosis, as revealed by increased NF-kB DNA binding activity in a time-dependent manner and attenuation of its proapoptotic effect by N-tosyl-L-phenylalanyl-chloromethyl ketone, an NF-kB blocker.

Conclusions

These data show that caspase-8 and -9 are involved in the apoptotic effect of celecoxib in cervical cancer cells. This requires the FADD-dependent pathway in a cell type-specific manner. In addition, NF-κB may play a key role in celecoxib-induced apoptosis.

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Abbreviations

COX:

Cyclooxygenase

NSAID:

Nonsteroidal anti-inflammatory drug

FADD:

Fas-associated death domain

TPCK:

N-tosyl-L-phenylalanyl-chloromethyl ketone

PGE2 :

Prostaglandin E2

EMSA:

Electrophoretic mobility shift assay

References

  1. Agarwal B, Rao CV, Bhendwal S, Ramey WR, Shirin H, Reddy BS, Holt PR (1999) Lovastatin augments sulindac-induced apoptosis in colon cancer cells and potentiates chemopreventive effects of sulindac. Gastroenterology 117:838–847

  2. Aragane Y, Kulms D, Metze D, Wilkes G, Poppelmann B, Luger TA, Schwarz T (1998) Ultraviolet light induces apoptosis via direct activation of CD95 (Fas/APO-1) independently of its ligand CD95L. J Cell Biol 140:171–182

  3. Bae SH, Jung ES, Park YM, Kim BS, Kim BK, Kim DG, Ryu WS (2001) Expression of cyclooxygenase-2 (COX-2) in hepatocellular carcinoma and growth inhibition of hepatoma cell lines by a COX-2 inhibitor, NS-398. Clin Cancer Res 7:1410–1418

  4. Boldin MP, Varfolomeev EE, Pancer Z, Mett IL, Camonis JH, Wallach D (1995) A novel protein that interacts with the death domain of Fas/APO1 contains a sequence motif related to the death domain. J Biol Chem 270:7795–7798

  5. 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 81:294–299

  6. Davies NM, McLachlan AJ, Day RO, Williams KM (2000) Clinical pharmacokinetics and pharmacodynamics of celecoxib: a selective cyclo-oxygenase-2 inhibitor. Clin Pharmacokinet 38:225–242

  7. Denkert C, Kobel M, Pest S, Koch I, Berger S, Schwabe M, Siegert A, Reles A, Klosterhalfen B, Hauptmann S (2002) Expression of cyclooxygenase 2 is an independent prognostic factor in human ovarian carcinoma. Am J Pathol 160:893–903

  8. Dubois RN (2000) Review article: cyclooxygenase—a target for colon cancer prevention. Aliment Pharmacol Ther 14[Suppl 1]:64–67

  9. Elder DJ, Paraskeva C (1998) COX-2 inhibitors for colorectal cancer. Nat Med 4:392–393

  10. Ferrandina G, Lauriola L, Distefano MG, Zannoni GF, Gessi M, Legge F, Maggiano N, Mancuso S, Capelli A, Scambia G, Ranelletti FO (2002a) Increased cyclooxygenase-2 expression is associated with chemotherapy resistance and poor survival in cervical cancer patients. J Clin Oncol 20:973–981

  11. Ferrandina G, Ranelletti FO, Lauriola L, Fanfani F, Legge F, Mottolese M, Nicotra MR, Natali PG, Zakut VH, Scambia G (2002b) Cyclooxygenase-2 (COX-2), epidermal growth factor receptor (EGFR), and Her-2/neu expression in ovarian cancer. Gynecol Oncol 85:305–310

  12. Gaffney DK, Holden J, Davis M, Zempolich K, Murphy KJ, Dodson M (2001) Elevated cyclooxygenase-2 expression correlates with diminished survival in carcinoma of the cervix treated with radiotherapy. Int J Radiat Oncol Biol Phys 49:1213–1217

  13. Goluboff ET, Shabsigh A, Saidi JA, Weinstein IB, Mitra N, Heitjan D, Piazza GA, Pamukcu R, Buttyan R, Olsson CA (1999) Exisulind (sulindac sulfone) suppresses growth of human prostate cancer in a nude mouse xenograft model by increasing apoptosis. Urology 53:440–445

  14. Grossman EM, Longo WE, Panesar N, Mazuski JE, Kaminski DL (2000) The role of cyclooxygenase enzymes in the growth of human gall bladder cancer cells. Carcinogenesis 21:1403–1409

  15. Han SS, Keum YS, Seo HJ, Chun KS, Lee SS, Surh YJ (2001a) Capsaicin suppresses phorbol ester-induced activation of NF-kappaB/Rel and AP-1 transcription factors in mouse epidermis. Cancer Lett 164:119–126

  16. Han Z, Pantazis P, Wyche JH, Kouttab N, Kidd VJ, Hendrickson EA (2001b) A Fas-associated death domain protein-dependent mechanism mediates the apoptotic action of non-steroidal anti-inflammatory drugs in the human leukemic Jurkat cell line. J Biol Chem 276:38748–38754

  17. Hengartner MO (2000) The biochemistry of apoptosis. Nature 407: 770-776

  18. Higashi Y, Kanekura T, Kanzaki T (2000) Enhanced expression of cyclooxygenase (COX)-2 in human skin epidermal cancer cells: evidence for growth suppression by inhibiting COX-2 expression. Int J Cancer 86:667–671

  19. Hsu AL, Ching TT, Wang DS, Song X, Rangnekar VM, Chen CS (2000a) The cyclooxygenase-2 inhibitor celecoxib induces apoptosis by blocking Akt activation in human prostate cancer cells independently of Bcl-2. J Biol Chem 275:11397–11403

  20. Hsu AL, Ching TT, Wang DS, Song X, Rangnekar VM, Chen CS (2000b) The cyclooxygenase-2 inhibitor celecoxib induces apoptosis by blocking Akt activation in human prostate cancer cells independently of Bcl-2. J Biol Chem 275:11397–11403

  21. Husain SS, Szabo IL, Tamawski AS (2002) NSAID inhibition of GI cancer growth: clinical implications and molecular mechanisms of action. Am J Gastroenterol 97:542–553

  22. Ishiko O, Sumi T, Yoshida H, Matsumoto Y, Honda K, Deguchi M, Yamada R, Ogita S (2001) Association between overexpression of cyclooxygenase-2 and suppression of apoptosis in advanced cancer of the uterine cervix after cyclic balloon-occluded arterial infusion. Oncol Rep 8:1259–1263

  23. Jendrossek V, Handrick R, Belka C (2003) Celecoxib activates a novel mitochondrial apoptosis signaling pathway. Faseb J 17:1547–1549

  24. Jiang XH, Lam SK, Lin MC, Jiang SH, Kung HF, Slosberg ED, Soh JW, Weinstein IB, Wong BC (2002) Novel target for induction of apoptosis by cyclo-oxygenase-2 inhibitor SC-236 through a protein kinase C-beta(1)-dependent pathway. Oncogene 21:6113–6122

  25. Johnson AJ, Song X, Hsu A, Chen C (2001) Apoptosis signaling pathways mediated by cyclooxygenase-2 inhibitors in prostate cancer cells. Adv Enzyme Regul 41:221–235

  26. Khuri FR, Wu H, Lee JJ, Kemp BL, Lotan R, Lippman SM, Feng L, Hong WK, Xu XC (2001) Cyclooxygenase-2 overexpression is a marker of poor prognosis in stage I non-small cell lung cancer. Clin Cancer Res 7:861–867

  27. Kim SG, Kim SN, Jong HS, Kim NK, Hong SH, Kim SJ, Bang YJ (2001) Caspase-mediated Cdk2 activation is a critical step to execute transforming growth factor-beta1-induced apoptosis in human gastric cancer cells. Oncogene 20:1254–1265

  28. Kulkarni S, Rader JS, Zhang F, Liapis H, Koki AT, Masferrer JL, Subbaramaiah K, Dannenberg AJ (2001) Cyclooxygenase-2 is overexpressed in human cervical cancer. Clin Cancer Res 7:429–434

  29. Leahy KM, Ornberg RL, Wang Y, Zweifel BS, Koki AT, Masferrer JL (2002) Cyclooxygenase-2 inhibition by celecoxib reduces proliferation and induces apoptosis in angiogenic endothelial cells in vivo. Cancer Res 62:625–631

  30. Li M, Wu X, Xu XC (2001) Induction of apoptosis in colon cancer cells by cyclooxygenase-2 inhibitor NS398 through a cytochrome c-dependent pathway. Clin Cancer Res 7:1010–1016

  31. Morris CD, Armstrong GR, Bigley G, Green H, Attwood SE (2001) Cyclooxygenase- 2 expression in the Barrett’s metaplasia-dysplasia- adenocarcinoma sequence. Am J Gastroenterol 96:990–996

  32. Nagata S (1997) Apoptosis by death factor. Cell 88:355–365

  33. Niederberger E, Tegeder I, Vetter G, Schmidtko A, Schmidt H, Euchenhofer C, Brautigam L, Grosch S, Geisslinger G (2001) Celecoxib loses its anti-inflammatory efficacy at high doses through activation of NF-kappaB. Faseb J 15:1622–1624

  34. Nzeako UC, Guicciardi ME, Yoon JH, Bronk SF, Gores GJ (2002) COX-2 inhibits Fas-mediated apoptosis in cholangiocarcinoma cells. Hepatology 35:552–559

  35. Paulson SK, Zhang JY, Breau AP, Hribar JD, Liu NW, Jessen SM, Lawal YM, Cogburn JN, Gresk CJ, Markos CS, Maziasz TJ, Schoenhard GL, Burton EG (2000) Pharmacokinetics, tissue distribution, metabolism, and excretion of celecoxib in rats. Drug Metab Dispos 28:514–521

  36. Phillips RK, Wallace MH, Lynch PM, Hawk E, Gordon GB, Saunders BP, Wakabayashi N, Shen Y, Zimmerman S, Godio L, Rodrigues-Bigas M, Su LK, Sherman J, Kelloff G, Levin B, Steinbach G (2002) A randomised, double blind, placebo controlled study of celecoxib, a selective cyclooxygenase 2 inhibitor, on duodenal polyposis in familial adenomatous polyposis. Gut 50:857–860

  37. Ristimaki A, Nieminen O, Saukkonen K, Hotakainen K, Nordling S, Haglund C (2001) Expression of cyclooxygenase-2 in human transitional cell carcinoma of the urinary bladder. Am J Pathol 158:849–853

  38. Ryu HS, Chang KH, Yang HW, Kim MS, Kwon HC, Oh KS (2000) High cyclooxygenase-2 expression in stage IB cervical cancer with lymph node metastasis or parametrial invasion. Gynecol Oncol 76:320–325

  39. Sales KJ, Katz AA, Davis M, Hinz S, Soeters RP, Hofmeyr MD, Millar RP, Jabbour HN (2001) Cyclooxygenase-2 expression and prostaglandin E(2) synthesis are up- regulated in carcinomas of the cervix: a possible autocrine/paracrine regulation of neoplastic cell function via EP2/EP4 receptors. J Clin Endocrinol Metab 86:2243–2249

  40. Schreinemachers DM, Everson RB (1994) Aspirin use and lung, colon, and breast cancer incidence in a prospective study. Epidemiology 5:138–146

  41. Shureiqi I, Chen D, Lotan R, Yang P, Newman RA, Fischer SM, Lippman SM (2000) 15-Lipoxygenase-1 mediates nonsteroidal anti-inflammatory drug-induced apoptosis independently of cyclooxygenase-2 in colon cancer cells. Cancer Res 60:6846–6850

  42. Smalley WE, DuBois RN (1997) Colorectal cancer and nonsteroidal anti-inflammatory drugs. Adv Pharmacol 39:1–20

  43. Song SH, Jong HS, Choi HH, Inoue H, Tanabe T, Kim NK, Bang YJ (2001) Transcriptional silencing of cyclooxygenase-2 by hyper-methylation of the 5’ CpG island in human gastric carcinoma cells. Cancer Res 61:4628–4635

  44. Song X, Lin HP, Johnson AJ, Tseng PH, Yang YT, Kulp SK, Chen CS (2002) Cyclooxygenase-2, player or spectator in cyclooxygenase-2 inhibitor-induced apoptosis in prostate cancer cells. J Natl Cancer Inst 94:585–591

  45. Soslow RA, Dannenberg AJ, Rush D, Woerner BM, Khan KN, Masferrer J, Koki AT (2000) COX-2 is expressed in human pulmonary, colonic, and mammary tumors. Cancer 89:2637–2645

  46. 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 15:1273–1275

  47. Steinbach G, Lynch PM, Phillips RK, Wallace MH, Hawk E, Gordon GB, Wakabayashi N, Saunders B, Shen Y, Fujimura T, Su LK, Levin B (2000) The effect of celecoxib, a cyclooxygenase-2 inhibitor, in familial adenomatous polyposis. N Engl J Med 342:1946–1952

  48. Suda T, Takahashi T, Golstein P, Nagata S (1993) Molecular cloning and expression of the Fas ligand, a novel member of the tumor necrosis factor family. Cell 75:1169–1178

  49. Tegeder I, Pfeilschifter J, Geisslinger G (2001) Cyclooxygenase-independent actions of cyclooxygenase inhibitors. Faseb J 15:2057–2072

  50. 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: 8021-8030

  51. Uotila P, Valve E, Martikainen P, Nevalainen M, Nurmi M, Harkonen P (2001) Increased expression of cyclooxygenase-2 and nitric oxide synthase-2 in human prostate cancer. Urol Res 29:23–28

  52. Vane JR, Bakhle YS, Botting RM (1998) Cyclooxygenases 1 and 2. Annu Rev Pharmacol Toxicol 38:97–120

  53. Wallach D, Varfolomeev EE, Malinin NL, Goltsev YV, Kovalenko AV, Boldin MP (1999) Tumor necrosis factor receptor and Fas signaling mechanisms. Annu Rev Immunol 17:331–367

  54. Waskewich C, Blumenthal RD, Li H, Stein R, Goldenberg DM, Burton J (2002) Celecoxib exhibits the greatest potency amongst cyclooxygenase (COX) inhibitors for growth inhibition of COX-2-negative hematopoietic and epithelial cell lines. Cancer Res 62:2029–2033

  55. Weppelmann B, Monkemeier D (1984) The influence of prostaglandin antagonists on radiation therapy of carcinoma of the cervix. Gynecol Oncol 17:196–199

  56. Williams CS, Mann M, DuBois RN (1999) The role of cyclooxygenases in inflammation, cancer, and development. Oncogene 18:7908–7916

  57. Wolf BB, Schuler M, Echeverri F, Green DR (1999) Caspase-3 is the primary activator of apoptotic DNA fragmentation via DNA fragmentation factor-45/inhibitor of caspase-activated DNase inactivation. J Biol Chem 274:30651–30656

  58. Wong BC, Zhu GH, Lam SK (1999) Aspirin induced apoptosis in gastric cancer cells. Biomed Pharmacother 53:315–318

  59. Zheng TS, Flavell RA (2000) Divinations and surprises: genetic analysis of caspase function in mice. Exp Cell Res 256:67–73

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Author information

Correspondence to Yong-Sang Song.

Additional information

This work was supported by a grant of the Korea Health 21R&D project, Ministry of Health & Welfare, Republic of Korea (01-PJ1-PG1-01CH05-0001) and Korean 2001–2003 BK 21 Project for Medicine, Dentistry & Pharmacy.

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Kim, S., Song, S., Kim, S. et al. Celecoxib induces apoptosis in cervical cancer cells independent of cyclooxygenase using NF-κB as a possible target. J Cancer Res Clin Oncol 130, 551–560 (2004). https://doi.org/10.1007/s00432-004-0567-6

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Keywords

  • COX-2
  • Celecoxib
  • Apoptosis
  • Caspase
  • FADD
  • NF-κB