Endocrinol Metab.  2014 Mar;29(1):54-61. 10.3803/EnM.2014.29.1.54.

Curcumin Enhances Docetaxel-Induced Apoptosis of 8505C Anaplastic Thyroid Carcinoma Cells

• Hong JM
• Park CS
• Nam-Goong
• Kim YS
• Lee JC
• Han MW
• Choi JI
• Kim YI
• Kim ES

• KMID: 2169352
• DOI: http://doi.org/10.3803/EnM.2014.29.1.54

Abstract

BACKGROUND
Anaplastic thyroid cancer (ATC) is one of the most aggressive malignancies in humans, and its progression is poorly controlled by existing therapeutic methods. Curcumin has been shown to suppress inflammation and angiogenesis. In this study, we evaluated whether curcumin could augment docetaxel-induced apoptosis of ATC cells. We also analyzed changes in nuclear factor kappaB (NF-kappaB) and cyclooxygenase-2 (COX-2) expression levels to delineate possible mechanisms of their combined action.
METHODS
ATC cells were cultured and treated with curcumin and docetaxel alone or in combination. The effects on cell viability were determined by MTS assay. Apoptosis was assessed by annexin V staining and confirmed by flow cytometric analysis. Caspase, COX-2, NF-kappaB levels were assayed by Western blotting.
RESULTS
Curcumin combined with docetaxel led to lower cell viability than treatment with docetaxel or curcumin alone. Annexin V staining followed by flow cytometric analysis demonstrated that curcumin treatment enhanced the docetaxel-induced apoptosis of ATC cells. Additionally, curcumin inhibited docetaxel-induced p65 activation and COX-2 expression.
CONCLUSION
We conclude that curcumin may enhance docetaxel's antitumor activity in ATC cells by interfering with NF-kappaB and COX-2. Our results suggest that curcumin may emerge as an attractive therapeutic candidate to enhance the antitumor activity of taxanes in ATC treatment.

Keyword

Thyroid cancer, anaplastic; Apoptosis; Curcumin; Cyclooxygenase 2; Docetaxel; NF-kappa B

MeSH Terms

Annexin A5
Apoptosis*
Blotting, Western
Cell Survival
Curcumin*
Cyclooxygenase 2
Humans
Inflammation
NF-kappa B
Taxoids
Thyroid Gland*
Thyroid Neoplasms*
Annexin A5
Curcumin
Cyclooxygenase 2
NF-kappa B
Taxoids

Figure

• 

  Fig. 1 Effects of docetaxel and/or curcumin treatment on the proliferation of 8505C, CAL62, FRO, and KTC2 cells. (A) 8505C cells were grown on 96-well plates and treated with varying doses of docetaxel (DOC) or curcumin (CUR) for 24 hours. (B) 8505C cells were grown on 96-well plates and treated with a combination of docetaxel (5 nM) and curcumin (10 µM) for 24 hours. (C) CAL62, FRO, and KTC2 cells were treated with docetaxel (5 nM) and curcumin (10 µM). Bars represent mean±SD. CTL, cell only; NS, no significant. aP<0.001; bP<0.05.

• 

  Fig. 2 Apoptosis-promoting effects of docetaxel (DOC) and curcumin (CUR) treatment. 8505C cells were cultured in 100-mm petri dishes and treated with docetaxel (5 nM) and/or curcumin (10 µM) for 24 hours. Apoptosis was assessed by Annexin V staining followed by flow cytometry analysis. Bars represent mean±SD. CTL, control.

• 

  Fig. 3 Effects of docetaxel (DOC) and curcumin (CUR) treatment on the expression of proapoptotic proteins. 8505C cells were treated with docetaxel (5 nM) and/or curcumin (10 µM) for 24 hours. Expression of cleaved caspase-3 and cleaved caspase-9 were detected by Western blot analysis. Actin was used as an internal control. Relative expression values are presented in the bar graph. Bars represent mean±SD. CTL, control.

• 

  Fig. 4 Effect of docetaxel (DOC) and curcumin (CUR) treatment on the nuclear factor κB (NF-κB) pathway. 8505C cells were treated with docetaxel (5 nM) and/or curcumin (10 µM) for 24 hours. Nuclear NF-κB was detected by Western blot analysis. Actin was used as control. Relative expression values are presented in the bar graph. Bars represent mean±SD. CTL, control.

• 

  Fig. 5 Effect of docetaxel (DOC) and curcumin (CUR) treatment on the expression of cyclooxygenase-2 (COX-2) protein. 8505C cells were treated with docetaxel (5 nM) and/or curcumin (10 µM) for 24 hours. Expression of COX-2 was detected by Western blot analysis. Actin was used as an internal control. Relative expression values are presented in the bar graph. Bars represent mean±SD. CTL, control.

Reference

1. Brignardello E, Gallo M, Baldi I, Palestini N, Piovesan A, Grossi E, Ciccone G, Boccuzzi G. Anaplastic thyroid carcinoma: clinical outcome of 30 consecutive patients referred to a single institution in the past 5 years. Eur J Endocrinol. 2007; 156:425–430.

2. Karunagaran D, Rashmi R, Kumar TR. Induction of apoptosis by curcumin and its implications for cancer therapy. Curr Cancer Drug Targets. 2005; 5:117–129.

3. Epstein J, Sanderson IR, Macdonald TT. Curcumin as a therapeutic agent: the evidence from in vitro, animal and human studies. Br J Nutr. 2010; 103:1545–1557.

4. Kawamori T, Lubet R, Steele VE, Kelloff GJ, Kaskey RB, Rao CV, Reddy BS. Chemopreventive effect of curcumin, a naturally occurring anti-inflammatory agent, during the promotion/progression stages of colon cancer. Cancer Res. 1999; 59:597–601.

5. Aggarwal BB, Shishodia S, Takada Y, Banerjee S, Newman RA, Bueso-Ramos CE, Price JE. Curcumin suppresses the paclitaxel-induced nuclear factor-kappaB pathway in breast cancer cells and inhibits lung metastasis of human breast cancer in nude mice. Clin Cancer Res. 2005; 11:7490–7498.

6. LoTempio MM, Veena MS, Steele HL, Ramamurthy B, Ramalingam TS, Cohen AN, Chakrabarti R, Srivatsan ES, Wang MB. Curcumin suppresses growth of head and neck squamous cell carcinoma. Clin Cancer Res. 2005; 11(19 Pt 1):6994–7002.

7. Cruz-Correa M, Shoskes DA, Sanchez P, Zhao R, Hylind LM, Wexner SD, Giardiello FM. Combination treatment with curcumin and quercetin of adenomas in familial adenomatous polyposis. Clin Gastroenterol Hepatol. 2006; 4:1035–1038.

8. Schiff PB, Fant J, Horwitz SB. Promotion of microtubule assembly in vitro by taxol. Nature. 1979; 277:665–667.

9. Yvon AM, Wadsworth P, Jordan MA. Taxol suppresses dynamics of individual microtubules in living human tumor cells. Mol Biol Cell. 1999; 10:947–959.

10. Mason KA, Hunter NR, Milas M, Abbruzzese JL, Milas L. Docetaxel enhances tumor radioresponse in vivo. Clin Cancer Res. 1997; 3(12 Pt 1):2431–2438.

11. Kim JC, Saha D, Cao Q, Choy H. Enhancement of radiation effects by combined docetaxel and flavopiridol treatment in lung cancer cells. Radiother Oncol. 2004; 71:213–221.

12. Reiner T, de las Pozas A, Gomez LA, Perez-Stable C. Low dose combinations of 2-methoxyestradiol and docetaxel block prostate cancer cells in mitosis and increase apoptosis. Cancer Lett. 2009; 276:21–31.

13. Tannock IF, de Wit R, Berry WR, Horti J, Pluzanska A, Chi KN, Oudard S, Theodore C, James ND, Turesson I, Rosenthal MA, Eisenberger MA. TAX 327 Investigators. Docetaxel plus prednisone or mitoxantrone plus prednisone for advanced prostate cancer. N Engl J Med. 2004; 351:1502–1512.

14. Kawada K, Kitagawa K, Kamei S, Inada M, Mitsuma A, Sawaki M, Kikumori T, Fujimoto Y, Arima H, Imai T, Ando Y. The feasibility study of docetaxel in patients with anaplastic thyroid cancer. Jpn J Clin Oncol. 2010; 40:596–599.

15. Meng Z, Mitsutake N, Nakashima M, Starenki D, Matsuse M, Takakura S, Namba H, Saenko V, Umezawa K, Ohtsuru A, Yamashita S. Dehydroxymethylepoxyquinomicin, a novel nuclear factor-kappaB inhibitor, enhances antitumor activity of taxanes in anaplastic thyroid cancer cells. Endocrinology. 2008; 149:5357–5365.

16. Troch M, Koperek O, Scheuba C, Dieckmann K, Hoffmann M, Niederle B, Raderer M. High efficacy of concomitant treatment of undifferentiated (anaplastic) thyroid cancer with radiation and docetaxel. J Clin Endocrinol Metab. 2010; 95:E54–E57.

17. Foote RL, Molina JR, Kasperbauer JL, Lloyd RV, McIver B, Morris JC, Grant CS, Thompson GB, Richards ML, Hay ID, Smallridge RC, Bible KC. Enhanced survival in locoregionally confined anaplastic thyroid carcinoma: a single-institution experience using aggressive multimodal therapy. Thyroid. 2011; 21:25–30.

18. Kurebayashi J, Otsuki T, Tanaka K, Yamamoto Y, Moriya T, Sonoo H. Medroxyprogesterone acetate decreases secretion of interleukin-6 and parathyroid hormone-related protein in a new anaplastic thyroid cancer cell line, KTC-2. Thyroid. 2003; 13:249–258.

19. Kojic SL, Strugnell SS, Wiseman SM. Anaplastic thyroid cancer: a comprehensive review of novel therapy. Expert Rev Anticancer Ther. 2011; 11:387–402.

20. Harris PJ, Bible KC. Emerging therapeutics for advanced thyroid malignancies: rationale and targeted approaches. Expert Opin Investig Drugs. 2011; 20:1357–1375.

21. Ain KB, Egorin MJ, DeSimone PA. Collaborative Anaplastic Thyroid Cancer Health Intervention Trials (CATCHIT) Group. Treatment of anaplastic thyroid carcinoma with paclitaxel: phase 2 trial using ninety-six-hour infusion. Thyroid. 2000; 10:587–594.

22. Higashiyama T, Ito Y, Hirokawa M, Fukushima M, Uruno T, Miya A, Matsuzuka F, Miyauchi A. Induction chemotherapy with weekly paclitaxel administration for anaplastic thyroid carcinoma. Thyroid. 2010; 20:7–14.

23. Barkett M, Gilmore TD. Control of apoptosis by Rel/NF-kappaB transcription factors. Oncogene. 1999; 18:6910–6924.

24. Lee R, Collins T. Nuclear factor-kappaB and cell survival: IAPs call for support. Circ Res. 2001; 88:262–264.

25. Zhang H, Morisaki T, Nakahara C, Matsunaga H, Sato N, Nagumo F, Tadano J, Katano M. PSK-mediated NF-kappaB inhibition augments docetaxel-induced apoptosis in human pancreatic cancer cells NOR-P1. Oncogene. 2003; 22:2088–2096.

26. Nakahara C, Nakamura K, Yamanaka N, Baba E, Wada M, Matsunaga H, Noshiro H, Tanaka M, Morisaki T, Katano M. Cyclosporin-A enhances docetaxel-induced apoptosis through inhibition of nuclear factor-kappaB activation in human gastric carcinoma cells. Clin Cancer Res. 2003; 9:5409–5416.

27. Domingo-Domenech J, Oliva C, Rovira A, Codony-Servat J, Bosch M, Filella X, Montagut C, Tapia M, Campas C, Dang L, Rolfe M, Ross JS, Gascon P, Albanell J, Mellado B. Interleukin 6, a nuclear factor-kappaB target, predicts resistance to docetaxel in hormone-independent prostate cancer and nuclear factor-kappaB inhibition by PS-1145 enhances docetaxel antitumor activity. Clin Cancer Res. 2006; 12:5578–5586.

28. Kikuchi E, Horiguchi Y, Nakashima J, Kuroda K, Oya M, Ohigashi T, Takahashi N, Shima Y, Umezawa K, Murai M. Suppression of hormone-refractory prostate cancer by a novel nuclear factor kappaB inhibitor in nude mice. Cancer Res. 2003; 63:107–110.

29. Shehzad A, Wahid F, Lee YS. Curcumin in cancer chemoprevention: molecular targets, pharmacokinetics, bioavailability, and clinical trials. Arch Pharm (Weinheim). 2010; 343:489–499.

30. Siwak DR, Shishodia S, Aggarwal BB, Kurzrock R. Curcumin-induced antiproliferative and proapoptotic effects in melanoma cells are associated with suppression of IkappaB kinase and nuclear factor kappaB activity and are independent of the B-Raf/mitogen-activated/extracellular signal-regulated protein kinase pathway and the Akt pathway. Cancer. 2005; 104:879–890.

31. Schaffer M, Schaffer PM, Zidan J, Bar Sela G. Curcuma as a functional food in the control of cancer and inflammation. Curr Opin Clin Nutr Metab Care. 2011; 14:588–597.

32. Soung YH, Chung J. Curcumin inhibition of the functional interaction between integrin alpha6beta4 and the epidermal growth factor receptor. Mol Cancer Ther. 2011; 10:883–891.

33. Lin SS, Lai KC, Hsu SC, Yang JS, Kuo CL, Lin JP, Ma YS, Wu CC, Chung JG. Curcumin inhibits the migration and invasion of human A549 lung cancer cells through the inhibition of matrix metalloproteinase-2 and -9 and vascular endothelial growth factor (VEGF). Cancer Lett. 2009; 285:127–133.

34. Bava SV, Sreekanth CN, Thulasidasan AK, Anto NP, Cheriyan VT, Puliyappadamba VT, Menon SG, Ravichandran SD, Anto RJ. Akt is upstream and MAPKs are downstream of NF-kappaB in paclitaxel-induced survival signaling events, which are down-regulated by curcumin contributing to their synergism. Int J Biochem Cell Biol. 2011; 43:331–341.

35. Kamat AM, Sethi G, Aggarwal BB. Curcumin potentiates the apoptotic effects of chemotherapeutic agents and cytokines through down-regulation of nuclear factor-kappaB and nuclear factor-kappaB-regulated gene products in IFN-alpha-sensitive and IFN-alpha-resistant human bladder cancer cells. Mol Cancer Ther. 2007; 6:1022–1030.

36. Merchan JR, Jayaram DR, Supko JG, He X, Bubley GJ, Sukhatme VP. Increased endothelial uptake of paclitaxel as a potential mechanism for its antiangiogenic effects: potentiation by Cox-2 inhibition. Int J Cancer. 2005; 113:490–498.

37. Tsujii M, Kawano S, Tsuji S, Sawaoka H, Hori M, DuBois RN. Cyclooxygenase regulates angiogenesis induced by colon cancer cells. Cell. 1998; 93:705–716.

38. McGinty A, Chang YW, Sorokin A, Bokemeyer D, Dunn MJ. Cyclooxygenase-2 expression inhibits trophic withdrawal apoptosis in nerve growth factor-differentiated PC12 cells. J Biol Chem. 2000; 275:12095–12101.

39. Ruggeri RM, Sciacchitano S, Vitarelli E, Trimarchi F, Barresi G, Trovato M. Immunoexpression of multidrug-resistance protein 2 and cyclooxygenase 2 in medullary thyroid carcinomas. Arch Pathol Lab Med. 2006; 130:1014–1019.

40. Cornetta AJ, Russell JP, Cunnane M, Keane WM, Rothstein JL. Cyclooxygenase-2 expression in human thyroid carcinoma and Hashimoto's thyroiditis. Laryngoscope. 2002; 112:238–242.