, 13:904 | Cite as

Coupling endoplasmic reticulum stress to the cell death program in mouse melanoma cells: effect of curcumin

  • Jason Bakhshi
  • Lee Weinstein
  • Karen S. Poksay
  • Brian Nishinaga
  • Dale E. Bredesen
  • Rammohan V. Rao
Original paper


The microenvironment of cancerous cells includes endoplasmic reticulum (ER) stress the resistance to which is required for the survival and growth of tumors. Acute ER stress triggers the induction of a family of ER stress proteins that promotes survival and/or growth of the cancer cells, and also confers resistance to radiation and chemotherapy. Prolonged or severe ER stress, however, may ultimately overwhelm the cellular protective mechanisms, triggering cell death through specific programmed cell death (pcd) pathways. Thus, downregulation of the protective stress proteins may offer a new therapeutic approach to cancer treatment. In this regard, recent reports have demonstrated the roles of the phytochemical curcumin in the inhibition of proteasomal activity and triggering the accumulation of cytosolic Ca2+ by inhibiting the Ca2+-ATPase pump, both of which enhance ER stress. Using a mouse melanoma cell line, we investigated the possibility that curcumin may trigger ER stress leading to programmed cell death. Our studies demonstrate that curcumin triggers ER stress and the activation of specific cell death pathways that feature caspase cleavage and activation, p23 cleavage, and downregulation of the anti-apoptotic Mcl-1 protein.


Endoplasmic reticulum Curcumin ER stress Caspase Apoptosis Programmed cell death 



Endoplasmic reticulum


Programmed cell death


Eukaryotic initiation factor-2 alpha


Glucose regulated protein



We thank members of the Bredesen laboratory for helpful comments and discussions and Molly Susag for administrative assistance. This work was supported by grants from the National Institutes of Health (NS33376 to D.E.B. & R.V.R, AG12282 and NS45093 to D.E.B) and Elisabeth R. Levy and Family Foundation award.


  1. 1.
    Rao RV, Ellerby HM, Bredesen DE (2004) Coupling endoplasmic reticulum stress to the cell death program. Cell Death Differ 11:372–380PubMedCrossRefGoogle Scholar
  2. 2.
    Rao RV, Bredesen DE (2004) Misfolded proteins, endoplasmic reticulum stress and neurodegeneration. Curr Opin Cell Biol 16:653–662PubMedCrossRefGoogle Scholar
  3. 3.
    Bredesen DE, Rao RV, Mehlen P (2006) Cell death in the nervous system. Nature 443:796–802PubMedCrossRefGoogle Scholar
  4. 4.
    Harding HP, Calfon M, Urano F, Novoa I, Ron D (2002) Transcriptional and translational control in the Mammalian unfolded protein response. Annu Rev Cell Dev Biol 18:575–599PubMedCrossRefGoogle Scholar
  5. 5.
    Rutkowski DT, Kaufman RJ (2004) A trip to the ER: coping with stress. Trends Cell Biol 14:20–28PubMedCrossRefGoogle Scholar
  6. 6.
    Lee AS (2001) The glucose-regulated proteins: stress induction and clinical applications. Trends Biochem Sci 26:504–510PubMedCrossRefGoogle Scholar
  7. 7.
    Rao RV, Castro-Obregon S, Frankowski H, Schuler M, Stoka V, Del Rio G, Bredesen DE, Ellerby HM (2002) Coupling endoplasmic reticulum stress to the cell death program. AN Apaf-1-INDEPENDENT INTRINSIC PATHWAY. J Biol Chem 277:21836–21842PubMedCrossRefGoogle Scholar
  8. 8.
    Rao RV, Peel A, Logvinova A, del Rio G, Hermel E, Yokota T, Goldsmith PC, Ellerby LM, Ellerby HM, Bredesen DE (2002) Coupling endoplasmic reticulum stress to the cell death program: role of the ER chaperone GRP78. FEBS Lett 514:122–128PubMedGoogle Scholar
  9. 9.
    Rao RV, Hermel E, Castro-Obregon S, del Rio G, Ellerby LM, Ellerby HM, Bredesen DE (2001) Coupling endoplasmic reticulum stress to the cell death program. Mechanism of caspase activation. J Biol Chem 276:33869–33874PubMedCrossRefGoogle Scholar
  10. 10.
    Lee AS, Hendershot LM (2006) ER stress and cancer. Cancer Biol Ther 5:721–722PubMedGoogle Scholar
  11. 11.
    Lee AS (2007) GRP78 induction in cancer: therapeutic and prognostic implications. Cancer Res 67:3496–3499PubMedCrossRefGoogle Scholar
  12. 12.
    Pae HO, Jeong SO, Jeong GS, Kim KM, Kim HS, Kim SA, Kim YC, Kang SD, Kim BN, Chung HT (2007) Curcumin induces pro-apoptotic endoplasmic reticulum stress in human leukemia HL-60 cells. Biochem Biophys Res Commun 353:1040–1045PubMedCrossRefGoogle Scholar
  13. 13.
    Bush JA, Cheung KJ Jr, Li G (2001) Curcumin induces apoptosis in human melanoma cells through a Fas receptor/caspase-8 pathway independent of p53. Exp Cell Res 271:305–314PubMedCrossRefGoogle Scholar
  14. 14.
    Surh YJ, Chun KS (2007) Cancer chemopreventive effects of curcumin. Adv Exp Med Biol 595:149–172PubMedGoogle Scholar
  15. 15.
    Aggarwal BB, Sundaram C, Malani N, Ichikawa H (2007) Curcumin: the Indian solid gold. Adv Exp Med Biol 595:1–75PubMedCrossRefGoogle Scholar
  16. 16.
    Shishodia S, Chaturvedi MM, Aggarwal BB (2007) Role of curcumin in cancer therapy. Curr Probl Cancer 31:243–305PubMedCrossRefGoogle Scholar
  17. 17.
    Jana NR, Dikshit P, Goswami A, Nukina N (2004) Inhibition of proteasomal function by curcumin induces apoptosis through mitochondrial pathway. J Biol Chem 279:11680–11685PubMedCrossRefGoogle Scholar
  18. 18.
    Logan-Smith MJ, East JM, Lee AG (2002) Evidence for a global inhibitor-induced conformation change on the Ca(2+)-ATPase of sarcoplasmic reticulum from paired inhibitor studies. Biochemistry 41:2869–2875PubMedCrossRefGoogle Scholar
  19. 19.
    Logan-Smith MJ, Lockyer PJ, East JM, Lee AG (2001) Curcumin, a molecule that inhibits the Ca2+-ATPase of sarcoplasmic reticulum but increases the rate of accumulation of Ca2+. J Biol Chem 276:46905–46911PubMedCrossRefGoogle Scholar
  20. 20.
    Ellerby HM, Martin SJ, Ellerby LM, Naiem SS, Rabizadeh S, Salvesen GS, Casiano CA, Cashman NR, Green DR, Bredesen DE (1997) Establishment of a cell-free system of neuronal apoptosis: comparison of premitochondrial, mitochondrial, and postmitochondrial phases. J Neurosci 17:6165–6178PubMedGoogle Scholar
  21. 21.
    Egger L, Madden DT, Rheme C, Rao RV, Bredesen DE (2007) Endoplasmic reticulum stress-induced cell death mediated by the proteasome. Cell Death Differ 14:1172–1180PubMedCrossRefGoogle Scholar
  22. 22.
    Rao RV, Poksay KS, Castro-Obregon S, Schilling B, Row RH, Del Rio G, Gibson BW, Ellerby HM, Bredesen DE (2004) Molecular components of a cell death pathway activated by endoplasmic reticulum stress. J Biol Chem 279:177–187PubMedCrossRefGoogle Scholar
  23. 23.
    Chinta SJ, Andersen JK (2006) Reversible inhibition of mitochondrial complex I activity following chronic dopaminergic glutathione depletion in vitro: implications for Parkinson’s disease. Free Radic Biol Med 41:1442–1448PubMedCrossRefGoogle Scholar
  24. 24.
    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
  25. 25.
    Liu H, Bowes RC III, van de Water B, Sillence C, Nagelkerke JF, Stevens JL (1997) Endoplasmic reticulum chaperones GRP78 and calreticulin prevent oxidative stress, Ca2+ disturbances, and cell death in renal epithelial cells. J Biol Chem 272:21751–21759PubMedCrossRefGoogle Scholar
  26. 26.
    Fu Y, Lee AS (2006) Glucose regulated proteins in cancer progression, drug resistance and immunotherapy. Cancer Biol Ther 5:741–744PubMedGoogle Scholar
  27. 27.
    Yamaguchi H, Wang HG (2004) CHOP is involved in endoplasmic reticulum stress-induced apoptosis by enhancing DR5 expression in human carcinoma cells. J Biol Chem 279:45495–45502PubMedCrossRefGoogle Scholar
  28. 28.
    Boyce M, Bryant KF, Jousse C, Long K, Harding HP, Scheuner D, Kaufman RJ, Ma D, Coen DM, Ron D, Yuan J (2005) A selective inhibitor of eIF2alpha dephosphorylation protects cells from ER stress. Science 307:935–939PubMedCrossRefGoogle Scholar
  29. 29.
    Sokka AL, Putkonen N, Mudo G, Pryazhnikov E, Reijonen S, Khiroug L, Belluardo N, Lindholm D, Korhonen L (2007) Endoplasmic reticulum stress inhibition protects against excitotoxic neuronal injury in the rat brain. J Neurosci 27:901–908PubMedCrossRefGoogle Scholar
  30. 30.
    Riedl SJ, Salvesen GS (2007) The apoptosome: signalling platform of cell death. Nat Rev Mol Cell Biol 8:405–413PubMedCrossRefGoogle Scholar
  31. 31.
    Tan TW, Tsai HR, Lu HF, Lin HL, Tsou MF, Lin YT, Tsai HY, Chen YF, Chung JG (2006) Curcumin-induced cell cycle arrest and apoptosis in human acute promyelocytic leukemia HL-60 cells via MMP changes and caspase-3 activation. Anticancer Res 26:4361–4371PubMedGoogle Scholar
  32. 32.
    Bae JH, Park JW, Kwon TK (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 303:1073–1079PubMedCrossRefGoogle Scholar
  33. 33.
    Nakano T, Watanabe H, Ozeki M, Asai M, Katoh H, Satoh H, Hayashi H (2006) Endoplasmic reticulum Ca2+ depletion induces endothelial cell apoptosis independently of caspase-12. Cardiovasc Res 69:908–915PubMedCrossRefGoogle Scholar
  34. 34.
    Bartlett JD, Luethy JD, Carlson SG, Sollott SJ, Holbrook NJ (1992) Calcium ionophore A23187 induces expression of the growth arrest and DNA damage inducible CCAAT/enhancer-binding protein (C/EBP)-related gene, gadd153. Ca2+ increases transcriptional activity and mRNA stability. J Biol Chem 267:20465–20470PubMedGoogle Scholar
  35. 35.
    Oyadomari S, Mori M (2004) Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ 11:381–389PubMedCrossRefGoogle Scholar
  36. 36.
    Lazebnik YA, Kaufmann SH, Desnoyers S, Poirier GG, Earnshaw WC (1994) Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature 371:346–347PubMedCrossRefGoogle Scholar
  37. 37.
    Rao RV, Niazi K, Mollahan P, Mao X, Crippen D, Poksay KS, Chen S, Bredesen DE (2006) Coupling endoplasmic reticulum stress to the cell-death program: a novel HSP90-independent role for the small chaperone protein p23. Cell Death Differ 13:415–425PubMedCrossRefGoogle Scholar
  38. 38.
    Germain M, Shore GC (2003) Cellular distribution of Bcl-2 family proteins. Sci STKE 2003:pe10PubMedGoogle Scholar
  39. 39.
    Reed JC (2006) Proapoptotic multidomain Bcl-2/Bax-family proteins: mechanisms, physiological roles, and therapeutic opportunities. Cell Death Differ 13:1378–1386PubMedCrossRefGoogle Scholar
  40. 40.
    Zong WX, Li C, Hatzivassiliou G, Lindsten T, Yu QC, Yuan J, Thompson CB (2003) Bax and Bak can localize to the endoplasmic reticulum to initiate apoptosis. J Cell Biol 162:59–69PubMedCrossRefGoogle Scholar
  41. 41.
    Scorrano L, Oakes SA, Opferman JT, Cheng EH, Sorcinelli MD, Pozzan T, Korsmeyer SJ (2003) BAX and BAK regulation of endoplasmic reticulum Ca2+: a control point for apoptosis. Science 300:135–139PubMedCrossRefGoogle Scholar
  42. 42.
    Yeh TC, Chiang PC, Li TK, Hsu JL, Lin CJ, Wang SW, Peng CY, Guh JH (2007) Genistein induces apoptosis in human hepatocellular carcinomas via interaction of endoplasmic reticulum stress and mitochondrial insult. Biochem Pharmacol 73:782–792PubMedCrossRefGoogle Scholar
  43. 43.
    Koshikawa N, Maejima C, Miyazaki K, Nakagawara A, Takenaga K (2006) Hypoxia selects for high-metastatic Lewis lung carcinoma cells overexpressing Mcl-1 and exhibiting reduced apoptotic potential in solid tumors. Oncogene 25:917–928PubMedCrossRefGoogle Scholar
  44. 44.
    Minagawa N, Kruglov EA, Dranoff JA, Robert ME, Gores GJ, Nathanson MH (2005) The anti-apoptotic protein Mcl-1 inhibits mitochondrial Ca2+ signals. J Biol Chem 280:33637–33644PubMedCrossRefGoogle Scholar
  45. 45.
    Subramanian T, Vijayalingam S, Lomonosova E, Zhao LJ, Chinnadurai G (2007) Evidence for involvement of BH3-only proapoptotic members in adenovirus-induced apoptosis. J Virol 81:10486–10495PubMedCrossRefGoogle Scholar
  46. 46.
    Chiang PC, Kung FL, Huang DM, Li TK, Fan JR, Pan SL, Shen YC, Guh JH (2006) Induction of Fas clustering and apoptosis by coral prostanoid in human hormone-resistant prostate cancer cells. Eur J Pharmacol 542:22–30PubMedCrossRefGoogle Scholar
  47. 47.
    Schuler M, Green DR (2001) Mechanisms of p53-dependent apoptosis. Biochem Soc Trans 29:684–688PubMedCrossRefGoogle Scholar
  48. 48.
    Liu FT, Newland AC, Jia L (2003) Bax conformational change is a crucial step for PUMA-mediated apoptosis in human leukemia. Biochem Biophys Res Commun 310:956–962PubMedCrossRefGoogle Scholar
  49. 49.
    Reimertz C, Kogel D, Rami A, Chittenden T, Prehn JH (2003) Gene expression during ER stress-induced apoptosis in neurons: induction of the BH3-only protein Bbc3/PUMA and activation of the mitochondrial apoptosis pathway. J Cell Biol 162:587–597PubMedCrossRefGoogle Scholar
  50. 50.
    Cartron PF, Gallenne T, Bougras G, Gautier F, Manero F, Vusio P, Meflah K, Vallette FM, Juin P (2004) The first alpha helix of Bax plays a necessary role in its ligand-induced activation by the BH3-only proteins Bid and PUMA. Mol Cell 16:807–818PubMedCrossRefGoogle Scholar
  51. 51.
    Schroder M, Kaufman RJ (2005) ER stress and the unfolded protein response. Mutat Res 569:29–63PubMedGoogle Scholar
  52. 52.
    Feldman DE, Chauhan V, Koong AC (2005) The unfolded protein response: a novel component of the hypoxic stress response in tumors. Mol Cancer Res 3:597–605PubMedCrossRefGoogle Scholar
  53. 53.
    Nakagawa T, Yuan J (2000) Cross-talk between two cysteine protease families. Activation of caspase-12 by calpain in apoptosis. J Cell Biol 150:887–894PubMedCrossRefGoogle Scholar
  54. 54.
    Nakagawa T, Zhu H, Morishima N, Li E, Xu J, Yankner BA, Yuan J (2000) Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-beta. Nature 403:98–103PubMedCrossRefGoogle Scholar
  55. 55.
    Hitomi J, Katayama T, Eguchi Y, Kudo T, Taniguchi M, Koyama Y, Manabe T, Yamagishi S, Bando Y, Imaizumi K, Tsujimoto Y, Tohyama M (2004) Involvement of caspase-4 in endoplasmic reticulum stress-induced apoptosis and A{beta}-induced cell death. J Cell Biol 165:347–356PubMedCrossRefGoogle Scholar
  56. 56.
    Hsu CH, Cheng AL (2007) Clinical studies with curcumin. Adv Exp Med Biol 595:471–480PubMedGoogle Scholar
  57. 57.
    Panchatcharam M, Miriyala S, Gayathri VS, Suguna L (2006) Curcumin improves wound healing by modulating collagen and decreasing reactive oxygen species. Mol Cell Biochem 290:87–96PubMedCrossRefGoogle Scholar
  58. 58.
    Huang MT, Ma W, Yen P, Xie JG, Han J, Frenkel K, Grunberger D, Conney AH (1997) Inhibitory effects of topical application of low doses of curcumin on 12-O-tetradecanoylphorbol-13-acetate-induced tumor promotion and oxidized DNA bases in mouse epidermis. Carcinogenesis 18:83–88PubMedCrossRefGoogle Scholar
  59. 59.
    Chun KS, Keum YS, Han SS, Song YS, Kim SH, Surh YJ (2003) Curcumin inhibits phorbol ester-induced expression of cyclooxygenase-2 in mouse skin through suppression of extracellular signal-regulated kinase activity and NF-kappaB activation. Carcinogenesis 24:1515–1524PubMedCrossRefGoogle Scholar
  60. 60.
    Huang MT, Newmark HL, Frenkel K (1997) Inhibitory effects of curcumin on tumorigenesis in mice. J Cell Biochem Suppl 27:26–34PubMedCrossRefGoogle Scholar
  61. 61.
    Cheng AL, Hsu CH, Lin JK, Hsu MM, Ho YF, Shen TS, Ko JY, Lin JT, Lin BR, Ming-Shiang W, Yu HS, Jee SH, Chen GS, Chen TM, Chen CA, Lai MK, Pu YS, Pan MH, Wang YJ, Tsai CC, Hsieh CY (2001) Phase I clinical trial of curcumin, a chemopreventive agent, in patients with high-risk or pre-malignant lesions. Anticancer Res 21:2895–2900PubMedGoogle Scholar
  62. 62.
    Cheng EH, Kirsch DG, Clem RJ, Ravi R, Kastan MB, Bedi A, Ueno K, Hardwick JM (1997) Conversion of Bcl-2 to a Bax-like death effector by caspases. Science 278:1966–1968PubMedCrossRefGoogle Scholar
  63. 63.
    Li H, Zhu H, Xu CJ, Yuan J (1998) Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. Cell 94:491–501PubMedCrossRefGoogle Scholar
  64. 64.
    Le Gouill S, Podar K, Harousseau JL, Anderson KC (2004) Mcl-1 regulation and its role in multiple myeloma. Cell Cycle 3:1259–1262PubMedGoogle Scholar
  65. 65.
    Zhang J, D’Ercole AJ (2004) Expression of Mcl-1 in cerebellar granule neurons is regulated by IGF-I in a developmentally specific fashion. Brain Res Dev Brain Res 152:255–263PubMedCrossRefGoogle Scholar
  66. 66.
    Bae J, Leo CP, Hsu SY, Hsueh AJ (2000) MCL-1S, a splicing variant of the antiapoptotic BCL-2 family member MCL-1, encodes a proapoptotic protein possessing only the BH3 domain. J Biol Chem 275:25255–25261PubMedCrossRefGoogle Scholar
  67. 67.
    Chae HJ, Kim HR, Xu C, Bailly-Maitre B, Krajewska M, Krajewski S, Banares S, Cui J, Digicaylioglu M, Ke N, Kitada S, Monosov E, Thomas M, Kress CL, Babendure JR, Tsien RY, Lipton SA, Reed JC (2004) BI-1 regulates an apoptosis pathway linked to endoplasmic reticulum stress. Mol Cell 15:355–366PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Jason Bakhshi
    • 1
  • Lee Weinstein
    • 2
  • Karen S. Poksay
    • 3
  • Brian Nishinaga
    • 4
  • Dale E. Bredesen
    • 3
    • 5
  • Rammohan V. Rao
    • 3
  1. 1.Terra Linda High SchoolSan RafaelUSA
  2. 2.Undergraduate Program, University of CaliforniaBerkeleyUSA
  3. 3.The Buck Institute for Age ResearchNovatoUSA
  4. 4.Albert Einstein College of MedicineBronxUSA
  5. 5.University of CaliforniaSan FranciscoUSA

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