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Curcumin induces apoptosis through the mitochondria-mediated apoptotic pathway in HT-29 cells

Journal of Zhejiang University SCIENCE B volume 10, pages 93–102 (2009)Cite this article

Abstract

Objective

To investigate the effects of curcumin on release of cytochrome c and expressions of Bcl-2, Bax, Bad, Bcl-xL, caspase-3, poly ADP-ribose polymerase (PARP), and survivin of HT-29 cells. Methods: HT-29 cells were treated with curcumin (0∼80 μmol/L) for 24 h. The release of cytochrome c from the mitochondria and the apoptosis-related proteins Bax, Bcl-2, Bcl-xL, Bad, caspase-3, PARP, and survivin were determined by Western blot analysis and their mRNA expressions by reverse transcriptase-polymerase chain reaction (RT-PCR).

Results

Curcumin significantly induced the growth inhibition and apoptosis of HT-29 cells. A decrease in expressions of Bcl-2, Bcl-xL and survivin was observed after exposure to 10∼80 μmol/L curcumin, while the levels of Bax and Bad increased in the curcumin-treated cells. Curcumin also induced the release of cytochrome c, the activation of caspase-3, and the cleavage of PARP in a dose-dependent manner.

Conclusion

These data suggest that curcumin induced the HT-29 cell apoptosis possibly via the mitochondria-mediated pathway.

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References

  • Aggarwal, S., Ichikawa, H., Takada, Y., Sandur, S.K., Shishodia, S., Aggarwal, B.B., 2006. Curcumin (diferuloylmethane) down-regulates expression of cell proliferation and antiapoptotic and metastatic gene products through suppression of IκBα kinase and Akt activation. Mol. Pharmacol., 69(1):195–206. [doi:10.1124/mol.105.017400]

    PubMed  CAS  Google Scholar 

  • Bae, J.H., Park, J.W., Kwon, T.K., 2003. Ruthenium red, inhibitor of mitochondrial Ca2+ uniporter, inhibits curcumin induced apoptosis via the prevention of intracellular Ca2+ depletion and cytochrome c release. Biochem. Biophys. Res. Commun., 303(4):1073–1079. [doi:10.1016/S0006-291X(03)00479-0]

    PubMed  Article  CAS  Google Scholar 

  • Bhaumik, S., Jyothi, M.D., Khar, A., 2000. Differential modulation of nitric oxide production by curcumin in host macrophages and NK cells. FEBS Lett., 483(1):78–82. [doi:10.1016/S0014-5793(00)02089-5]

    PubMed  Article  CAS  Google Scholar 

  • Cao, J., Liu, Y., Jia, L., Zhou, H.M., Kong, Y., Yang, G., Jiang, L.P., Li, Q.J., Zhong, L.F., 2007. Curcumin induces apoptosis through mitochondrial hyperpolarization and mtDNA damage in human hepatoma G2 cells. Free Radic. Biol. Med., 43(6):968–975. [doi:10.1016/j.freeradbiomed.2007.06.006]

    PubMed  Article  CAS  Google Scholar 

  • Chadalapaka, G., Jutooru, I., Chintharlapalli, S., Papineni, S., Smith, R., Li, X., Safe, S., 2008. Curcumin decreases specificity protein expression in bladder cancer cells. Cancer Res., 68(13):5345–5354. [doi:10.1158/0008-5472.CAN-07-6805]

    PubMed  Article  CAS  Google Scholar 

  • Chen, C.Y., Liu, T.Z., Liu, Y.W., Tseng, W.C., Liu, R.H., Lu, F.J., Lin, Y.S., Chen, C.H., 2007. 6-Shogaol (alkanone from Ginger) induces apoptotic cell death of human hepatoma p53 mutant Mahlavu subline via an oxidative stress-mediated caspase-dependent mechanism. J. Agric. Food Chem., 55(3):948–954. [doi:10.1021/jf0624594]

    PubMed  Article  CAS  Google Scholar 

  • Chen, J.H., Cao, J.L., Chu, Y.L., Wang, Z.L., Yang, Z.T., Wang, H.L., 2008. T-2 toxin-induced apoptosis involving Fas, p53, Bcl-xL, Bcl-2, Bax and caspase-3 signaling pathways in human chondrocytes. J. Zhejiang Univ. Sci. B, 9(6):455–463. [doi:10.1631/jzus.B0820013]

    PubMed  Article  CAS  Google Scholar 

  • Cory, S., Adams, J.M., 2002. The Bcl-2 family: regulators of the cellular life-or-death switch. Nat. Rev. Cancer, 2(9):647–656. [doi:10.1038/nrc883]

    PubMed  Article  CAS  Google Scholar 

  • Desai, B.N., Myers, B.R., Schreiber, S.L., 2002. FKBP12-rapamycin-associated protein associates with mitochondria and senses osmotic stress via mitochondrial dysfunction. Proc. Natl. Acad. Sci. USA, 99(7):4319–4324. [doi:10.1073/pnas.261702698]

    PubMed  Article  CAS  Google Scholar 

  • Gao, X., Kuo, J., Jiang, H., Deeb, D., Liu, Y., Divine, G., Chapman, R.A., Dulchavsky, S.A., Gautam, S.C., 2004. Immunomodulatory activity of curcumin: suppression of lymphocyte proliferation, development of cell-mediated cytotoxicity, and cytokine production in vitro. Biochem. Pharmacol., 68(1):51–61. [doi:10.1016/j.bcp.2004.03.015]

    PubMed  Article  CAS  Google Scholar 

  • Green, D.R., 2000. Apoptotic pathways: paper wraps stone blunts scissors. Cell, 102(1):1–4. [doi:10.1016/S0092-8674(00)00003-9]

    PubMed  Article  CAS  Google Scholar 

  • Grishko, V., Rachek, L., Musiyenko, S., Ledoux, S.P., Wilson, G.L., 2005. Involvement of mtDNA damage in free fatty acid-induced apoptosis. Free Radic. Biol. Med., 38(6):755–762. [doi:10.1016/j.freeradbiomed.2004.11.023]

    PubMed  Article  CAS  Google Scholar 

  • Khor, T.O., Gul, Y.A., Ithnin, H., Seow, H.F., 2006. A comparative study of the expression of Wnt-1, WISP-1, survivin and cyclin-D1 in colorectal carcinoma. Int. J. Colorectal Dis., 21(4):291–300. [doi:10.1007/s00384-005-0002-8]

    PubMed  Article  Google Scholar 

  • Kluck, R.M., Bossy-Wetzel, E., Green, D.R., Newmeyer, D.D., 1997. The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science, 275(5303):1132–1136. [doi:10.1126/science.275.5303.1132]

    PubMed  Article  CAS  Google Scholar 

  • Lam, A.K., Saleh, S., Smith, R.A., Ho, Y., 2008. Quantitative analysis of survivin in colorectal adenocarcinoma: increased expression and correlation with telomerase activity. Human Pathol., 39(8):1229–1233. [doi:10.1016/j.humpath.2008.01.001]

    Article  CAS  Google Scholar 

  • Li, F., Brattain, M.G., 2006. Role of the survivin gene in pathophysiology. Am. J. Pathol., 169(1):1–10. [doi:10.2353/ajpath.2006.060121]

    PubMed  Article  CAS  Google Scholar 

  • Li, P., Nijhawan, D., Budihardjo, I., Srinivasula, S.M., Ahmad, M., Alnemri, E.S., Wang, X., 1997. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell, 91(4):479–489. [doi:10.1016/S0092-8674(00)80434-1]

    PubMed  Article  CAS  Google Scholar 

  • Moiseeva, E.P., Almeida, G.M., Jones, G.D.D., Manson, M.M., 2007. Extended treatment with physiologic concentrations of dietary phytochemicals results in altered gene expression, reduced growth, and apoptosis of cancer cells. Mol. Cancer Ther., 6(11):3071–3079. [doi:10.1158/1535-7163.MCT-07-0117]

    PubMed  Article  CAS  Google Scholar 

  • Newman, D.J., Cragg, G.M., Snader, K.M., 2003. Natural products as sources of new drugs over the period 1981–2002. J. Nat. Prod., 66(7):1022–1037. [doi:10. 1021/np030096l]

    PubMed  Article  CAS  Google Scholar 

  • Pan, M.H., Chang, W.L., Lin-Shiau, S.Y., Ho, C.T., Lin, J.K., 2001. Induction of apoptosis by garcinol and curcumin through cytochrome c release and activation of caspases in human leukemia HL-60 cells. J. Agric. Food Chem., 49(3):1464–1474. [doi:10.1021/jf001129v]

    PubMed  Article  CAS  Google Scholar 

  • Piwocka, K., Zabłocki, K., Wieckowski, M.R., Skierski, J., Feiga, I., Szopa, J., Drela, N., Wojtczak, L., Sikora, E., 1999. A novel apoptosis-like pathway, independent of mitochondria and caspases, induced by curcumin in human lymphoblastoid T (Jurkat) cells. Exp. Cell Res., 249(2):299–307. [doi:10.1006/excr.1999.4480]

    PubMed  Article  CAS  Google Scholar 

  • Porn-Ares, M.I., Chow, S.C., Slotte, J.P., Orrenius, S., 1997. Induction of apoptosis and potentiation of TNF and Fas-mediated apoptosis in U937 cells by the xanthogenate compound D609. Exp. Cell Res., 235(1):48–54. [doi:10.1006/excr.1997.3641]

    PubMed  Article  CAS  Google Scholar 

  • Rashmi, R., Santhosh Kumar, T.R., Karunagaran, D., 2003. Human colon cancer cells differ in their sensitivity to curcumin-induced apoptosis and heat shock protects them by inhibiting the release of apoptosis-inducing factor and caspases. FEBS Lett., 538(1–3):19–24. [doi:10.1016/S0014-5793(03)00099-1]

    PubMed  Article  CAS  Google Scholar 

  • Reed, J.C., 1998. Bcl-2 family proteins. Oncogene, 17(25):3225–3236. [doi:10.1038/sj.onc.1202591]

    PubMed  Article  Google Scholar 

  • Salvioli, S., Ardizzoni, A., Franceschi, C., Cossarizza, A., 1997. JC-1, but not DiOC6(3) or rhodamine 123, is a reliable fluorescent probe to assess Δϕ changes in intact cells: implications for studies on mitochondrial functionality during apoptosis. FEBS Lett., 411(1):77–82. [doi:10.1016/S0014-5793(97)00669-8]

    PubMed  Article  CAS  Google Scholar 

  • Scott, D.W., Loo, G., 2004. Curcumin-induced GADD153 gene up-regulation in human colon cancer cells. Carcinogenesis, 25(11):2155–2164. [doi:10.1093/carcin/ bgh239]

    PubMed  Article  CAS  Google Scholar 

  • Sen, S., Sharma, H., Singh, N., 2005. Curcumin enhances Vinorelbine-mediated apoptosis in NSCLC cells by the mitochondrial pathway. Biochem. Biophys. Res. Commun., 331(4):1245–1252. [doi:10.1016/j.bbrc.2005.04. 044]

    PubMed  Article  CAS  Google Scholar 

  • Shang, T., Joseph, J., Hillard, C.J., Kalyanaraman, B., 2005. Death-associated protein kinase as a sensor of mitochondrial membrane potential: role of lysosome in mitochondrial toxin-induced cell death. J. Biol. Chem., 280(41):34644–34653. [doi:10.1074/jbc.M506466200]

    PubMed  Article  CAS  Google Scholar 

  • Sharma, R.A., Gescher, A.J., Steward, W.P., 2005. Curcumin: the story so far. Eur. J. Cancer, 41(13):1955–1968. [doi:10.1016/j.ejca.2005.05.009]

    PubMed  Article  CAS  Google Scholar 

  • Shi, M., Cai, Q., Yao, L., Mao, Y., Ming, Y., Ouyang, G., 2006. Antiproliferation and apoptosis induced by curcumin in human ovarian cancer cells. Cell Biol. Internat., 30(3):221–226. [doi:10.1016/j.cellbi.2005.10.024]

    Article  CAS  Google Scholar 

  • Shishodia, S., Amin, H.M., Lai, R., Aggarwal, B.B., 2005a. Curcumin (diferuloylmethane) inhibits constitutive NF-κB activation, induces G1/S arrest, suppresses proliferation, and induces apoptosis in mantle cell lymphoma. Biochem. Pharmacol., 70(5):700–713. [doi:10. 1016/j.bcp.2005.04.043]

    PubMed  Article  CAS  Google Scholar 

  • Shishodia, S., Sethi, G., Aggarwal, B.B., 2005b. Curcumin: getting back to the roots. Ann. N. Y. Acad. Sci., 1056(1):206–217. [doi:10.1196/annals.1352.010]

    PubMed  Article  CAS  Google Scholar 

  • Song, G., Mao, Y.B., Cai, Q.F., Yao, L.M., Ouyang, G.L., Bao, D., 2005. Curcumin induces human HT-29 colon adenocarcinoma cell apoptosis by activating p53 and regulating apoptosis-related protein expression. Braz. J. Med. Biol. Res., 38(12):1791–1798. [doi:10.1590/S0100-879X2005001200007]

    PubMed  Article  CAS  Google Scholar 

  • Suzuki, A., Ito, T., Hayashida, M., Hayasaki, Y., Tuutomi, Y., Akahane, K., Nakano, T., Miura, M., Shiraki, K., 2000. Survivin initiates procaspase-3/p21 complex formation as a result of interaction with Cdk4 to resist Fas-mediated cell death. Oncogene, 19(10):1346–1353. [doi:10.1038/ sj.onc.1203429]

    PubMed  Article  CAS  Google Scholar 

  • Tan, H.Y., Liu, J., Wu, S.M., Luo, H.S., 2005. Expression of a novel apoptosis inhibitor-survivin in colorectal carcinoma. World J. Gastroenterol., 11(30):4689–4692.

    PubMed  CAS  Google Scholar 

  • Wang, X., 2001. The expanding role of mitochondria in apoptosis. Genes Dev., 15(22):2922–2933.

    PubMed  CAS  Google Scholar 

  • Watson, A.J.M., 2006. An overview of apoptosis and the prevention of colorectal cancer. Crit. Rev. Oncol. Hematol., 57(2):107–121. [doi:10.1016/j.critrevonc.2005.06. 005]

    PubMed  Article  Google Scholar 

  • Woo, J.H., Kim, Y.H., Choi, Y.J., Kim, D.G., Lee, K.S., Bae, J.H., Mindo, S., Chang, J.S., Jeong, Y.J., Lee, Y.H., et al., 2003. Molecular mechanisms of curcumin-induced cytotoxicity: induction of apoptosis through generation of reactive oxygen species, down-regulation of Bcl-xL and IAP, the release of cytochrome c and inhibition of Akt. Carcinogenesis, 24(7):1199–1208. [doi:10.1093/carcin/ bgg082]

    PubMed  Article  CAS  Google Scholar 

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Affiliations

  1. Department of Chemistry, College of Science, Zhejiang University, Hangzhou, 310027, China

    Jin-bo Wang & Tian-xing Wu

  2. Ningbo Institute of Technology, Zhejiang University, Ningbo, 315100, China

    Jin-bo Wang, Li-li Qi & Shui-di Zheng

Authors
  1. Jin-bo Wang
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  2. Li-li Qi
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  3. Shui-di Zheng
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  4. Tian-xing Wu
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Corresponding author

Correspondence to Tian-xing Wu.

Additional information

Project (No. 2006C12098) supported by the Science and Technology Department of Zhejiang Province, China

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Wang, Jb., Qi, Ll., Zheng, Sd. et al. Curcumin induces apoptosis through the mitochondria-mediated apoptotic pathway in HT-29 cells. J. Zhejiang Univ. Sci. B 10, 93–102 (2009). https://doi.org/10.1631/jzus.B0820238

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  • DOI: https://doi.org/10.1631/jzus.B0820238

Key words

  • Curcumin
  • Apoptosis
  • Mitochondrial pathway
  • HT-29 cells

CLC number

  • Q81