Journal of Physiology and Biochemistry

, Volume 69, Issue 2, pp 289–296 | Cite as

Effects of curcumin on bleomycin-induced apoptosis in human malignant testicular germ cells

  • Aysegul Cort
  • Mujgan Timur
  • Evrim Ozdemir
  • Tomris Ozben
Original Paper


Testicular cancer is the most common cancer among young men of reproductive age. Bleomycin is a frequently used drug for the treatment of several malignancies and is part of the chemotherapy protocols in testicular cancer. Bleomycin causes an increase in oxidative stress which has been shown to induce apoptosis in cancer cells. Curcumin (diferuloylmethane), an active component of the spice turmeric, has attracted interest because of its anti-inflammatory and chemopreventive activities. However, no study has been carried out so far to elucidate its interaction with bleomycin in testicular cancer cells. In this study, we investigated the effects of curcumin and bleomycin on apoptosis signalling pathways and compared the effects of bleomycin with H2O2 which directly produces reactive oxygen species. We measured apoptosis markers such as caspase-3, caspase-8, and caspase-9 activities and Bcl-2, Bax, and Cyt-c levels in NCCIT cells incubated with curcumin (5 μM), bleomycin (120 μg/ml), bleomycin + curcumin, H2O2 (35 μM), and H2O2 + curcumin for 72 h. Curcumin, bleomycin, and H2O2 caused apoptosis indicated as increases in caspase-3, caspase-8, and caspase-9 activities and Bax and cytoplasmic Cyt-c levels and a decrease in Bcl-2 level. Concurrent use of curcumin with bleomycin decreased caspase activities and Bax and Cyt-c levels compared to their separate effects in NCCIT cells. Our findings suggest that concurrent use of curcumin during chemotherapy in testis cancer should be avoided due to the inhibitory effect of curcumin on bleomycin-induced apoptosis.


Testicular cancer NCCIT cell Apoptosis Bleomycin Curcumin 



This study was supported by Scientific and Technological Research Council, Turkey (TUBITAK) (COST-CM0603-15) and Akdeniz University.

Conflict of interest

The authors report no conflicts of interest. The content is solely the responsibility of the authors.


  1. 1.
    Aggarwal BB, Kumar A, Bharti AC (2003) Anticancer potential of curcumin: preclinical and clinical studies. Anticancer Res 23:363–398PubMedGoogle Scholar
  2. 2.
    Bejarano I, Espino J, Marchena AM, Barriga C, Paredes SD, Rodriguez AB, Pariente JA (2011) Melatonin enhances hydrogen peroxide-induced apoptosis in human promyelocytic leukaemia HL-60 cells. Mol Cell Biochem 353:167–176PubMedCrossRefGoogle Scholar
  3. 3.
    Chen QY, Lu GH, Wu YQ, Zheng Y, Xu K, Wu LJ, Jiang ZY, Feng R, Zhou JY (2010) Curcumin induces mitochondria pathway mediated cell apoptosis in A549 lung adenocarcinoma cells. Oncol Rep 23:1285–1292PubMedGoogle Scholar
  4. 4.
    Chen X, Zhong Z, Xu Z, Chen L, Wang Y (2011) No protective effect of curcumin on hydrogen peroxide-induced cytotoxicity in HepG2 cells. Pharmacol Rep 63:724–732PubMedGoogle Scholar
  5. 5.
    Choudhuri T, Pal S, Das T, Sa G (2005) Curcumin selectively induces apoptosis in deregulated cyclin D1-expressed cells at G2 phase of cell cycle in a p53-dependent manner. J Biol Chem 280:20059–20068PubMedCrossRefGoogle Scholar
  6. 6.
    Cort A, Timur M, Ozdemir E, Kucuksayan E, Ozben T (2012) Synergistic anticancer activity of curcumin and bleomycin: an in vitro study using human malignant testicular germ cells. Mol Med Report 5:1481–1486Google Scholar
  7. 7.
    Datta K, Babbar P, Srivastava T, Sinha S, Chattopadhyay P (2002) p53 dependent apoptosis in glioma cell lines in response to hydrogen peroxide induced oxidative stress. Int J Biochem Cell Biol 34:148–157PubMedCrossRefGoogle Scholar
  8. 8.
    de Wit R, Stoter G, Kaye SB, Sleijfer DT, Jones WG, ten Bokkel Huinink WW, Rea LA, Collette L, Sylvester R (1997) Importance of bleomycin in combination chemotherapy for good-prognosis testicular nonseminoma: a randomized study of the European Organization for Research and Treatment of Cancer Genitourinary Tract Cancer Cooperative Group. J Clin Oncol 15:1837–1843PubMedGoogle Scholar
  9. 9.
    Garcea G, Berry DP, Jones DJ, Singh R, Dennison AR, Farmer PB, Sharma RA, Steward WP, Gescher AJ (2005) Consumption of the putative chemopreventive agent curcumin by cancer patients: assessment of curcumin levels in the colorectum and their pharmacodynamic consequences. Cancer Epidemiol Biomarkers Prev 14:120–125PubMedGoogle Scholar
  10. 10.
    Hanif R, Qiao L, Shiff SJ, Rigas B (1997) Curcumin, a natural plant phenolic food additive, inhibits cell proliferation and induces cell cycle changes in colon adenocarcinoma cell lines by a prostaglandin-independent pathway. J Lab Clin Med 130:576–584PubMedCrossRefGoogle Scholar
  11. 11.
    Huang AC, Chang CL, Yu CS, Chen PY, Yang JS, Ji BC, Lin TP, Chiu CF, Yeh SP, Huang YP, Lien JC, Chung JG (2011) Induction of apoptosis by curcumin in murine myelomonocytic leukemia WEHI-3 cells is mediated via endoplasmic reticulum stress and mitochondria-dependent pathways. Environ Toxicol. doi: 10.1002/tox.20716
  12. 12.
    Huyghe E, Matsuda T, Thonneau P (2003) Increasing incidence of testicular cancer worldwide: a review. J Urol 170:5–11PubMedCrossRefGoogle Scholar
  13. 13.
    Ip SW, Wu SY, Yu CC, Kuo CL, Yu CS, Yang JS, Lin ZP, Chiou SM, Chung HK, Ho HC, Chung JG (2011) Induction of apoptotic death by curcumin in human tongue squamous cell carcinoma SCC-4 cells is mediated through endoplasmic reticulum stress and mitochondria-dependent pathways. Cell Biochem Funct 29:641–650PubMedCrossRefGoogle Scholar
  14. 14.
    Jiang MC, Yang-Yen HF, Yen JJ, Lin JK (1996) Curcumin induces apoptosis in immortalized NIH 3T3 and malignant cancer cell lines. Nutr Cancer 26:111–120PubMedCrossRefGoogle Scholar
  15. 15.
    Kim KY, Yu SN, Lee SY, Chun SS, Choi YL, Park YM, Song CS, Chatterjee B, Ahn SC (2011) Salinomycin-induced apoptosis of human prostate cancer cells due to accumulated reactive oxygen species and mitochondrial membrane depolarization. Biochem Biophys Res Commun 413:80–86PubMedCrossRefGoogle Scholar
  16. 16.
    Kuo ML, Huang TS, Lin JK (1996) Curcumin, an antioxidant and anti-tumor promoter, induces apoptosis in human leukemia cells. Biochim Biophys Acta 1317:95–100PubMedCrossRefGoogle Scholar
  17. 17.
    Lee JW, Hong HM, Kwon DD, Pae HO, Jeong HJ (2010) Dimethoxycurcumin, a structural analogue of curcumin, induces apoptosis in human renal carcinoma caki cells through the production of reactive oxygen species, the release of cytochrome C, and the activation of caspase-3. Kor J Urol 51:870–878CrossRefGoogle Scholar
  18. 18.
    Lin SS, Lai KC, Hsu SC, Yang JS, Kuo CL, Lin JP, Ma YS, Wu CC, Chung JG (2009) 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 285:127–133PubMedCrossRefGoogle Scholar
  19. 19.
    Liu E, Wu J, Cao W, Zhang J, Liu W, Jiang X, Zhang X (2007) Curcumin induces G2/M cell cycle arrest in a p53-dependent manner and upregulates ING4 expression in human glioma. J Neurooncol 85:263–270PubMedCrossRefGoogle Scholar
  20. 20.
    Llambi F, Green DR (2011) Apoptosis and oncogenesis: give and take in the BCL-2 family. Curr Opin Genet Dev 21:12–20PubMedCrossRefGoogle Scholar
  21. 21.
    Ozben T (2007) Oxidative stress and apoptosis: impact on cancer therapy. J Pharm Sci 96:2181–2196PubMedCrossRefGoogle Scholar
  22. 22.
    Punithavathi D, Venkatesan N, Babu M (2000) Curcumin inhibition of bleomycin-induced pulmonary fibrosis in rats. Br J Pharmacol 131:169–172PubMedCrossRefGoogle Scholar
  23. 23.
    Ramachandran C, You W (1999) Differential sensitivity of human mammary epithelial and breast carcinoma cell lines to curcumin. Breast Cancer Res Treat 54:269–278PubMedCrossRefGoogle Scholar
  24. 24.
    Rao J, Xu DR, Zheng FM, Long ZJ, Huang SS, Wu X, Zhou WH, Huang RW, Liu Q (2011) Curcumin reduces expression of Bcl-2, leading to apoptosis in daunorubicin-insensitive CD34+ acute myeloid leukemia cell lines and primary sorted CD34+ acute myeloid leukemia cells. J Transl Med 9:71PubMedCrossRefGoogle Scholar
  25. 25.
    Sharma RA, Ireson CR, Verschoyle RD, Hill KA, Williams ML, Leuratti C, Manson MM, Marnett LJ, Steward WP, Gescher A (2001) Effects of dietary curcumin on glutathione S-transferase and malondialdehyde-DNA adducts in rat liver and colon mucosa: relationship with drug levels. Clin Cancer Res 7:1452–1458PubMedGoogle Scholar
  26. 26.
    Shehzad A, Wahid F, Lee YS (2010) Curcumin in cancer chemoprevention: molecular targets, pharmacokinetics, bioavailability, and clinical trials. Arch Pharm (Weinheim) 343:489–499CrossRefGoogle Scholar
  27. 27.
    Sikora E, Bielak-Zmijewska A, Piwocka K, Skierski J, Radziszewska E (1997) Inhibition of proliferation and apoptosis of human and rat T lymphocytes by curcumin, a curry pigment. Biochem Pharmacol 54:899–907PubMedCrossRefGoogle Scholar
  28. 28.
    Somasundaram S, Edmund NA, Moore DT, Small GW, Shi YY, Orlowski RZ (2002) Dietary curcumin inhibits chemotherapy-induced apoptosis in models of human breast cancer. Cancer Res 62:3868–3875PubMedGoogle Scholar
  29. 29.
    Tourkina E, Gooz P, Oates JC, Ludwicka-Bradley A, Silver RM, Hoffman S (2004) Curcumin-induced apoptosis in scleroderma lung fibroblasts: role of protein kinase cepsilon. Am J Respir Cell Mol Biol 31:28–35PubMedCrossRefGoogle Scholar
  30. 30.
    Ulbright TM (1993) Germ cell neoplasms of the testis. Am J Surg Pathol 17:1075–1091PubMedCrossRefGoogle Scholar
  31. 31.
    Venkatesan N (1998) Curcumin attenuation of acute adriamycin myocardial toxicity in rats. Br J Pharmacol 124:425–427PubMedCrossRefGoogle Scholar
  32. 32.
    Venkatesan N, Punithavathi V, Chandrakasan G (1997) Curcumin protects bleomycin-induced lung injury in rats. Life Sci 61:PL51–PL58PubMedCrossRefGoogle Scholar
  33. 33.
    Weir NM, Selvendiran K, Kutala VK, Tong L, Vishwanath S, Rajaram M, Tridandapani S, Anant S, Kuppusamy P (2007) Curcumin induces G2/M arrest and apoptosis in cisplatin-resistant human ovarian cancer cells by modulating Akt and p38 MAPK. Cancer Biol Ther 6:178–184PubMedCrossRefGoogle Scholar
  34. 34.
    Wu SH, Hang LW, Yang JS, Chen HY, Lin HY, Chiang JH, Lu CC, Yang JL, Lai TY, Ko YC, Chung JG (2010) Curcumin induces apoptosis in human non-small cell lung cancer NCI-H460 cells through ER stress and caspase cascade- and mitochondria-dependent pathways. Anticancer Res 30:2125–2133PubMedGoogle Scholar
  35. 35.
    Yallapu MM, Maher DM, Sundram V, Bell MC, Jaggi M, Chauhan SC (2010) Curcumin induces chemo/radio-sensitization in ovarian cancer cells and curcumin nanoparticles inhibit ovarian cancer cell growth. J Ovarian Res 3:11PubMedCrossRefGoogle Scholar
  36. 36.
    Yang CL, Ma YG, Xue YX, Liu YY, Xie H, Qiu GR (2012) Curcumin induces small cell lung cancer NCI-H446 cell apoptosis via the reactive oxygen species-mediated mitochondrial pathway and not the cell death receptor pathway. DNA Cell Biol 31:139–150PubMedCrossRefGoogle Scholar
  37. 37.
    Yu J, Zhou X, He X, Dai M, Zhang Q (2011) Curcumin induces apoptosis involving bax/bcl-2 in human hepatoma SMMC-7721 cells. Asian Pac J Cancer Prev 12:1925–1929PubMedGoogle Scholar
  38. 38.
    Zhang X, Zhang HQ, Zhu GH, Wang YH, Yu XC, Zhu XB, Liang G, Xiao J, Li XK (2012) A novel mono-carbonyl analogue of curcumin induces apoptosis in ovarian carcinoma cells via endoplasmic reticulum stress and reactive oxygen species production. Mol Med Report 5:739–744Google Scholar
  39. 39.
    Zuliani T, Denis V, Noblesse E, Schnebert S, Andre P, Dumas M, Ratinaud MH (2005) Hydrogen peroxide-induced cell death in normal human keratinocytes is differentiation dependent. Free Radic Biol Med 38:307–316PubMedCrossRefGoogle Scholar

Copyright information

© University of Navarra 2012

Authors and Affiliations

  • Aysegul Cort
    • 1
  • Mujgan Timur
    • 1
  • Evrim Ozdemir
    • 1
  • Tomris Ozben
    • 1
  1. 1.Department of Biochemistry, Faculty of MedicineAkdeniz UniversityAntalyaTurkey

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