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Molecular and Cellular Biochemistry

, Volume 273, Issue 1–2, pp 57–67 | Cite as

Mechanism of inhibition of ascites tumor growth in mice by curcumin is mediated by NF-kB and caspase activated DNase

  • Madesh Belakavadi
  • Bharathi P. SalimathEmail author
Article

Abstract

One of the most clinically relevant biological activities of curcumin is its anti-cancer property, implicating multiple intracellular pathways in the process. In the present report, we investigated the effect of curcumin on the activation of apoptotic and anti-angiogenic pathways in Ehrlich Ascites Tumor (EAT) cells. Treatment with curcumin in vivo resulted in inhibition of proliferation of EAT cells and ascites formation. Further, we demonstrate that the induction of apoptosis in EAT cells showed nuclear condensation, DNA fragmentation and translocation of caspase-activated DNase (CAD) to nucleus upon curcumin treatment. Curcumin-induced apoptosis is mediated through activation of caspase-3, which is specifically inhibited by the caspase-3 inhibitor, Ac-DEVD-CHO. On the other hand, the decreased secretion of ascites by EAT cells is corroborated by reduction in VEGF secretion upon curcumin treatment. Further, CD31 immunohistological staining of peritoneum sections in curcumin-treated mice suggests its efficacy in acting as anti-angiogenic compound in EAT cells by inhibiting proliferation of endothelial cells in mouse peritoneum. However, immunoflurescence studies of NF-kB revealed that the inhibition of nuclear translocation of NF-kB p65, a transcription factor required for VEGF gene expression, in curcumin-treated EAT cells. These results suggest a further possible clinical application of this diet-derived compound curcumin, as both proapoptotic and anti-angiogenic compound in association with conventional chemotherapeutic agents.

Keywords

anti-angiogenesis apoptosis curcumin CAD VEGF NF-kB 

Abbreviations

VEGF

vascular endothelial cell growth factor

CAD

caspase activated DNase

NF-kB

Nuclear Factor kB

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References

  1. 1.
    Folkman J: Angiogenesis in cancer, vascular, rheumatoid and other diseases. Nat Med 1: 27–31, 1995PubMedGoogle Scholar
  2. 2.
    Hanahan D, Folkman J: Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell 86: 353–364, 1996CrossRefPubMedGoogle Scholar
  3. 3.
    Brem S: Angiogenesis and cancer control: From concept to therapeutic trials. Cancer Control 6: 436–458, 1999PubMedGoogle Scholar
  4. 4.
    Boehm T, Folkman J, Browder T, O’ Reilly MS: Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance. Nature 390: 404–407, 1997CrossRefPubMedGoogle Scholar
  5. 5.
    Pluda JM: Tumor-associated angiogenesis: Mechanisms, clinical implications, and therapeutic strategies. Semin Oncol 24: 203–218, 1997PubMedGoogle Scholar
  6. 6.
    Rak J, Kerbel RS: Treating cancer by inhibiting angiogenesis: New hopes and potential pitfalls. Cancer Metastasis Rev 15: 231–236, 1996CrossRefPubMedGoogle Scholar
  7. 7.
    Taraboletti G, Margosio B: Antiangiogenic and antivascular therapy for cancer. Curr Opin Pharmacol 1: 378–384, 2001CrossRefPubMedGoogle Scholar
  8. 8.
    Cao Y: Endogenous angiogenesis inhibitors and their therapeutic implications. Int J Biochem Cell Biol 33: 357–369, 2001CrossRefPubMedGoogle Scholar
  9. 9.
    Min JK, Han KY, Kim EC, Kim YM, Lee SW, Kim OH, Kim KW, Gho YS, Kwon YG: Capsaicin inhibits in vitro and in vivo angiogenesis. Cancer Res 64: 644–651, 2004PubMedGoogle Scholar
  10. 10.
    You YJ, Nam NH, Kim Y, Bae KH, Ahn BZ: Antiangiogenic activity of lupeol from Bombax ceiba. Phytother Res 17: 341–344, 2003CrossRefPubMedGoogle Scholar
  11. 11.
    Chow LM, Chui CH, Tang JC, Lau FY, Yau MY, Cheng GY, Wong RS, Lai PB, Leung TW, Teo IT, Cheung F, Guo D, Chan AS: Antiangiogenic potential of Gleditsia sinensis fruit extract. Int J Mol Med 12: 269–273, 2003PubMedGoogle Scholar
  12. 12.
    Sharma RA, McLelland HR, Hill KA, Ireson CR, Euden SA, Manson MM, Pirmohamed M, Marnett LJ, Gescher AJ, Steward WP: Pharmacodynamic and pharmacokinetic study of oral Curcuma extract in patients with colorectal cancer. Clin Cancer Res 7: 1894–1900, 2001PubMedGoogle Scholar
  13. 13.
    Stoner GD, Mukhtar H: Polyphenols as cancer chemopreventive agents. J Cell Biochem 22: 169–180, 1995PubMedGoogle Scholar
  14. 14.
    Nandkarni KM: Curcuma longa, Popular Prakashan, Bombay, 1976.Google Scholar
  15. 15.
    Govindaraja VS: Turmeric: Chemistry, technology, and quality. Crit Rev Food Sci Nutr 12: 199, 1980PubMedGoogle Scholar
  16. 16.
    Ammon HP, Wahl MA: Pharmacology of Curcuma longa. Planta Medica 57: 1–7, 1991PubMedGoogle Scholar
  17. 17.
    Huang MT, Want ZY, Georgiadis CA, Laskin JD, Conney AH: Inhibitory effects of curcumin on tumor initiation by benzo[a]pyrene and 7,12-dimethylbenz[a] anthracene. Carcinogenesis 13: 2183–2186, 1992PubMedGoogle Scholar
  18. 18.
    Azuine MA, Bhide SV: Chemopreventive effect of turmeric against stomach and skin tumors induced by chemical carcinogens in Swiss mice. Nutr Cancer 17: 77–83, 1992PubMedGoogle Scholar
  19. 19.
    Nagbhusan M, Bhide SV: Curcumin as an inhibitor of cancer. J Am Coll Nutr 11: 192–198, 1992PubMedGoogle Scholar
  20. 20.
    Huang MT, Smart RC, Wong C-Q, Conney AH: Inhibitory effect of curcumin, chlorogenic acid, caffeic acid, and ferulic acid on tumor promotion in mouse skin by 12-O-tetradecanoylphorbol-13-acetate. Cancer Res 48: 5941–5946, 1988PubMedGoogle Scholar
  21. 21.
    Conney AH, Lysz T, Ferraro T, Abidi TF, Manchand PS, Laskin JD, Huang MT: Inhibitory effect of curcumin and some related dietary compounds on tumor promotion and arachidonic acid metabolism in mouse skin. Adv Enzyme Regulation 31: 385–396, 1991CrossRefGoogle Scholar
  22. 22.
    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 59: 597–601, 1999PubMedGoogle Scholar
  23. 23.
    Liu JY, Lin SJ, Lin SK: Inhibitory effects of curcumin on protein kinase C activity induced by 12-O-tetradecanoyl-phorbol-13-acetate in NIH 3T3 cells. Carcinogenesis 14: 857–861, 1993PubMedGoogle Scholar
  24. 24.
    Pal S, Choudhuri T, Chattopadhyay S, Bhattacharya A, Datta G, Das T, Sa G: Mechanisms of curcumin-induced apoptosis of Ehrlich’s ascites carcinoma cells. Biochem Biophys Res Commun 288: 658–665, 2001CrossRefPubMedGoogle Scholar
  25. 25.
    Jee SH, Shen SC, Tseng CR, Chiu HC, Kuo ML: Curcumin induces a p53-dependent apoptosis in human basal cell carcinoma cells. J Invest Dermatol 111: 656–661, 1998CrossRefPubMedGoogle Scholar
  26. 26.
    Pan M, Lin-Shiau S, Lin J: Comparative studies on the suppression of nitric oxide synthase by curcumin and its hydrogenated metabolites through down-regulation of IkappaB kinase and NFkappaB activation in macrophages. Biochem Pharmacol 60: 1665–1676, 2000CrossRefPubMedGoogle Scholar
  27. 27.
    Korutla L, Kumar R: Inhibitory effect of curcumin on epidermal growth factor receptor kinase activity in A431 cells. Biochim Biophys Acta 1224: 597–600, 1994CrossRefPubMedGoogle Scholar
  28. 28.
    Singh S, Aggarwal BB: Activation of transcription factor NF-kappa B is suppressed by curcumin (diferuloylmethane). J Biol Chem 270: 24995–25000, 1995CrossRefPubMedGoogle Scholar
  29. 29.
    Jiang MC, Yang-Yen HF, Lin JK, Yen JJ: Differential regulation of p53, c-Myc, Bcl-2 and Bax protein expression during apoptosis induced by widely divergent stimuli in human hepatoblastoma cells. Oncogene 13: 609–616, 1996PubMedGoogle Scholar
  30. 30.
    Alfranca A, Gutierrez MD, Vara A, Aragones J, Vidal F, Landazuri MO: c-Jun and hypoxia-inducible factor 1 functionally cooperate in hypoxia-induced gene transcription. Mol Cell Biol 22: 12–22, 2002CrossRefPubMedGoogle Scholar
  31. 31.
    Shibata A, Nagaya T, Imai T, Funahashi H, Nakao A, Seo H: Inhibition of NF-kappa B activity decreases the VEGF mRNA expression in MDA-MB-231 breast cancer cells. Breast Cancer Res Treat 73: 237–243, 2002CrossRefPubMedGoogle Scholar
  32. 32.
    Thaloor D, Singh AK, Sidhu GS, Prasad PV, Kleinman HK, Maheswari RK: Inhibition of angiogenic differentiation of human umbilical vein endothelial cells by curcumin. Cell Growth Differ 9: 305–312, 1998PubMedGoogle Scholar
  33. 33.
    Mohan R, Sivak J, Ashton P, Russo LA, Pham BQ, Kasahara N, Raizman MB, Fini ME: Curcuminoids inhibit the angiogenic response stimulated by fibroblast growth factor-2, including expression of matrix metalloproteinase gelatinase B. J Biol Chem 275: 10405–10412, 2000CrossRefPubMedGoogle Scholar
  34. 34.
    Shim JS, Kim JH, Cho HY, Yum YN, Kim SH, Park HJ, Shim BS, Choi SH, Kwon HJ: Irreversible inhibition of CD13/aminopeptidase N by the antiangiogenic agent curcumin. Chem Biol 8: 695–704, 2003CrossRefGoogle Scholar
  35. 35.
    Shao ZM, Shen ZZ, Liu CH, Sartippour MR, Go VL, Heber D, Nguyen M: Curcumin exerts multiple suppressive effects on human breast carcinoma cells. Int J Cancer 98: 234–240, 2002CrossRefPubMedGoogle Scholar
  36. 36.
    Arbiser JL, Klauber N, van Rohan R, Leeuwen R, Huang MT, Fisher C, Flynn E, Byers HR: Curcumin is an in vivo inhibitor of angiogenesis. Mol Med 4: 376–383, 1998PubMedGoogle Scholar
  37. 37.
    Gururaj AE, Belakavadi M, Venkatesh DA, Marme D, Salimath BP: Molecular mechanisms of anti-angiogenic effect of curcumin. Biochem Biophys Res Commun 297: 934–942, 2002CrossRefPubMedGoogle Scholar
  38. 38.
    Srinivas G, John Anto R, Srinivas P, Vidhyalakshmi S, Priya Senan V, Karunagaran D: Emodin induces apoptosis of human cervical cancer cells through poly (ADP-ribose) polymerase cleavage and activation of caspase-9. Euro J Pharmacol 473: 117–125, 2003CrossRefGoogle Scholar
  39. 39.
    Keifer JA, Guttridge DC, Ashbumer BP, Baldwin AS Jr.: Inhibition of NF-kappa B activity by thalidomide through suppression of I kappa B kinase activity. J Biol Chem 276: 22382–22387, 2001PubMedGoogle Scholar
  40. 40.
    Kapahi P, Takahashi T, Natoli G, Adams SR, Chen Y, Tsien RY, Karin M: Inhibition of NF-kappa B activation by arsenite through reaction with a critical cysteine in the activation loop of I kappa B kinase. J Bid Chem 275: 36062–36066, 2000Google Scholar
  41. 41.
    Surh YJ, Han SS, Keum YS, Seo HJ, Lee SS: Inhibitory effects of curcumin and capsaicin on phorbol ester-induced activation of eukaryotic transcription factors, NF-kappa B and AP-1. Biofactors 12: 107–112, 2000PubMedGoogle Scholar
  42. 42.
    Huang MT, Lou YR, Ma M, Newmark HL, Reuhl KR, Conney AR: Inhibitory effects of dietary curcumin on fore stomach, duodenal, and colon carcinogenesis in mice. Cancer Res 54: 5841–5847, 1994PubMedGoogle Scholar
  43. 43.
    Rao CV, Rivenson A, Simi B, Reddy BS: Chemoprevention of colon carcinogenesis by dietary curcumin, a naturally occurring plant phenolic compound. Cancer Res 55: 259–266, 1995PubMedGoogle Scholar
  44. 44.
    Jiang M-C, Yang-Yen HF, Yen JJ, Lin JK: Curcumin induces apoptosis in immortalized NIH 3T3 and malignant cancer cell lines. Nutr Cancer 26: 111–120, 1996PubMedGoogle Scholar
  45. 45.
    Eamshaw WC, Martins LM, Kaufmann SH: Mammalian caspases: Structure, activation, substrates, and functions during apoptosis. Annu Rev Biochem 68: 383–424, 1999PubMedGoogle Scholar
  46. 46.
    Thornberry N, Lazebnik Y: Caspases: Enemies within. Science 281: 1312–1316, 1998PubMedGoogle Scholar
  47. 47.
    Widlak P: The DFF40/CAD endonuclease and its role in apoptosis. Acta Biochim Pol 47: 1037–1044, 2000PubMedGoogle Scholar
  48. 48.
    Weidner N, Carroll PR, Flax J, Bluementhal W, Folkman J: Tumor angiogenesis correlates with metastasis in invasive prostate cancer. Am J Pathol 143: 401–409, 1993PubMedGoogle Scholar
  49. 49.
    Shibuya M: Role of VEGF-FLT receptor system in normal and tumor angiogenesis. Adv Cancer Res 67: 281–316, 1995PubMedGoogle Scholar
  50. 50.
    Thaloor D, Singh AK, Sidhu GS, Prasad PV, Kleinman H, Maheswari RK: Inhibition of angiogenic differentiation of human umbilical vein endothelial cells by curcumin. Cell Growth Differ 9: 305–312, 1998PubMedGoogle Scholar
  51. 51.
    Perkins ND: The Rel/NF/kB family: Friend and Foe. Trends Biochem Sci 25: 434–440, 2000PubMedGoogle Scholar
  52. 52.
    Baldwin AS, Jr.: The NF-kB and IkB proteins: New discoveries and insights. Annu Rev Immunol 14: 649–683, 1996PubMedGoogle Scholar
  53. 53.
    Verma IM, Stevenson J: IkB kinase: Beginning not the end. Proc Natl Acad Sci USA, 94: 11758–11760, 1997PubMedGoogle Scholar
  54. 54.
    Gutteridge DC, Albanese C, Reuther JY, Pestell RG, Baldwin AS, Jr.: NF-kB controls cell growth and differentiation through transcriptional regulation of cyclin Dl. Mol Cell Biol 19: 5785–5799, 1999PubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  1. 1.Department of Applied Botany and BiotechnologyUniversity of MysoreIndia
  2. 2.Department of Applied Botany and BiotechnologyUniversity of MysoreMysoreIndia

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