Investigational New Drugs

, Volume 29, Issue 6, pp 1276–1283 | Cite as

The anti-cancer activity of dihydroartemisinin is associated with induction of iron-dependent endoplasmic reticulum stress in colorectal carcinoma HCT116 cells



Dihydroartemisinin (DHA), the main active metabolite of artemisinin derivatives, is among the artemisinin derivatives possessing potent anti-malarial and anti-cancer activities. In the present study, we found that DHA displayed significant anti-proliferative activity in human colorectal carcinoma HCT116 cells, which may be attributed to its induction of G1 phase arrest and apoptosis. To further elucidate the mechanism of action of DHA, a proteomic study employed two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was performed. Glucose-regulated protein 78 (GRP78), which is related with endoplasmic reticulum stress (ER stress), was identified to be significantly up-regulated after DHA treatment. Further study demonstrated that DHA enhanced expression of GRP78 as well as growth arrest and DNA-damage-inducible gene 153 (GADD153, another ER stress-associated molecule) at both mRNA and protein levels. DHA treatment also led to accumulation of GADD153 in cell nucleus. Moreover, pretreatment of HCT116 cells with the iron chelator deferoxamine mesylate salt (DFO) abrogated induction of GRP78 and GADD153 upon DHA treatment, indicating iron is required for DHA-induced ER stress. This result is consistent with the fact that the anti-proliferative activity of DHA is also mediated by iron. We thus suggest the unbalance of redox may result in DHA-induced ER stress, which may contribute, at least in part, to its anti-cancer activity.


Dihydroartemisinin 2-DE ER stress GRP78 GADD153 



Artemisinin and its derivatives


Two-dimensional gel electrophoresis


Deferoxamine mesylate salt




Endoplasmic reticulum


Growth arrest and DNA-damage-inducible gene 153


Glucose-regulated protein 78


Heat-shock proteins


Isoelectric focusing


Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry


Propidium iodide


Sulforhodamine B


  1. 1.
    White NJ (2008) Qinghaosu (artemisinin): the price of success. Science 320:330–334PubMedCrossRefGoogle Scholar
  2. 2.
    Lu JJ, Meng LH, Cai YJ et al (2008) Dihydroartemisinin induces apoptosis in HL-60 leukemia cells dependent of iron and p38 mitogen-activated protein kinase activation but independent of reactive oxygen species. Cancer Biol Ther 7:1017–1023PubMedCrossRefGoogle Scholar
  3. 3.
    Efferth T, Sauerbrey A, Olbrich A, Gebhart E, Rauch P, Weber HO et al (2003) Molecular modes of action of artesunate in tumor cell lines. Mol Pharmacol 64:382–394PubMedCrossRefGoogle Scholar
  4. 4.
    Efferth T, Olbrich A, Bauer R (2002) mRNA expression profiles for the response of human tumor cell lines to the antimalarial drugs artesunate, arteether, and artemether. Biochem Pharmacol 64:617–623PubMedCrossRefGoogle Scholar
  5. 5.
    Li LN, Zhang HD, Yuan SJ et al (2007) Artesunate attenuates the growth of human colorectal carcinoma and inhibits hyperactive Wnt/beta-catenin pathway. Int J Cancer 121:1360–1365PubMedCrossRefGoogle Scholar
  6. 6.
    Jiao Y, Ge CM, Meng QH et al (2007) Dihydroartemisinin is an inhibitor of ovarian cancer cell growth. Acta Pharmacol Sin 28:1045–1056PubMedCrossRefGoogle Scholar
  7. 7.
    Chen T, Li M, Zhang R et al (2009) Dihydroartemisinin induces apoptosis and sensitizes human ovarian cancer cells to carboplatin therapy. J Cell Mol Med 13:1358–1370PubMedCrossRefGoogle Scholar
  8. 8.
    Lu YY, Chen TS, Qu JL et al (2009) Dihydroartemisinin (DHA) induces caspase-3-dependent apoptosis in human lung adenocarcinoma ASTC-a-1 cells. J Biomed Sci 16:16PubMedCrossRefGoogle Scholar
  9. 9.
    Lu JJ, Meng LH, Shankavaram UT et al (2010) Dihydroartemisinin accelerates c-MYC oncoprotein degradation and induces apoptosis in c-MYC-overexpressing tumor cells. Biochem Pharmacol 80:22–30PubMedCrossRefGoogle Scholar
  10. 10.
    Chen HH, Zhou HJ, Wang WQ et al (2004) Antimalarial dihydroartemisinin also inhibits angiogenesis. Cancer Chemother Pharmacol 53:423–432PubMedCrossRefGoogle Scholar
  11. 11.
    Dell’Eva R, Pfeffer U, Vene R et al (2004) Inhibition of angiogenesis in vivo and growth of Kaposi’s sarcoma xenograft tumors by the anti-malarial artesunate. Biochem Pharmacol 68:2359–2366PubMedCrossRefGoogle Scholar
  12. 12.
    Mercer AE, Maggs JL, Sun XM et al (2007) Evidence for the involvement of carbon-centered radicals in the induction of apoptotic cell death by artemisinin compounds. J Biol Chem 282:9372–9382PubMedCrossRefGoogle Scholar
  13. 13.
    Efferth T, Benakis A, Romero MR et al (2004) Enhancement of cytotoxicity of artemisinins toward cancer cells by ferrous iron. Free Radic Biol Med 37:998–1009PubMedCrossRefGoogle Scholar
  14. 14.
    Efferth T (2006) Molecular pharmacology and pharmacogenomics of artemisinin and its derivatives in cancer cells. Curr Drug Targets 7:407–421PubMedCrossRefGoogle Scholar
  15. 15.
    Singh NP, Lai H (2001) Selective toxicity of dihydroartemisinin and holotransferrin toward human breast cancer cells. Life Sci 70:49–56PubMedCrossRefGoogle Scholar
  16. 16.
    Efferth T, Oesch F (2004) Oxidative stress response of tumor cells: microarray-based comparison between artemisinins and anthracyclines. Biochem Pharmacol 68:3–10PubMedCrossRefGoogle Scholar
  17. 17.
    Tao Z, Zhou Y, Lu J et al (2007) Caspase-8 preferentially senses the apoptosis-inducing action of NG-18, a Gambogic acid derivative, in human leukemia HL-60 cells. Cancer Biol Ther 6:691–696PubMedCrossRefGoogle Scholar
  18. 18.
    Yue QX, Cao ZW, Guan SH et al (2008) Proteomics characterization of the cytotoxicity mechanism of ganoderic acid D and computer-automated estimation of the possible drug target network. Mol Cell Proteomics 7:949–961PubMedCrossRefGoogle Scholar
  19. 19.
    Jiang XS, Tang LY, Cao XJ et al (2005) Two-dimensional gel electrophoresis maps of the proteome and phosphoproteome of primitively cultured rat mesangial cells. Electrophoresis 26:4540–4562PubMedCrossRefGoogle Scholar
  20. 20.
    Lee AS (1992) Mammalian stress response: induction of the glucose-regulated protein family. Curr Opin Cell Biol 4:267–273PubMedCrossRefGoogle Scholar
  21. 21.
    Wang XZ, Lawson B, Brewer JW et al (1996) Signals from the stressed endoplasmic reticulum induce C/EBP-homologous protein (CHOP/GADD153). Mol Cell Biol 16:4273–4280PubMedGoogle Scholar
  22. 22.
    Chen H, Sun B, Pan S et al (2009) Dihydroartemisinin inhibits growth of pancreatic cancer cells in vitro and in vivo. Anticancer Drugs 20:131–140PubMedCrossRefGoogle Scholar
  23. 23.
    Milli A, Cecconi D, Campostrini N et al (2008) A proteomic approach for evaluating the cell response to a novel histone deacetylase inhibitor in colon cancer cells. Biochim Biophys Acta 1784:1702–1710PubMedGoogle Scholar
  24. 24.
    Disbrow GL, Baege AC, Kierpiec KA et al (2005) Dihydroartemisinin is cytotoxic to papillomavirus-expressing epithelial cells in vitro and in vivo. Cancer Res 65:10854–10861PubMedCrossRefGoogle Scholar
  25. 25.
    Mu D, Zhang W, Chu D et al (2008) The role of calcium, P38 MAPK in dihydroartemisinin-induced apoptosis of lung cancer PC-14 cells. Cancer Chemother Pharmacol 61:639–645PubMedCrossRefGoogle Scholar
  26. 26.
    Kim I, Xu W, Reed JC (2008) Cell death and endoplasmic reticulum stress: disease relevance and therapeutic opportunities. Nat Rev Drug Discov 7:1013–1030PubMedCrossRefGoogle Scholar
  27. 27.
    Stockwin LH, Han B, Yu SX et al (2009) Artemisinin dimer anticancer activity correlates with heme-catalyzed reactive oxygen species generation and endoplasmic reticulum stress induction. Int J Cancer 125:1266–1275PubMedCrossRefGoogle Scholar
  28. 28.
    Brostrom CO, Brostrom MA (1998) Regulation of translational initiation during cellular responses to stress. Prog Nucleic Acid Res Mol Biol 58:79–125PubMedCrossRefGoogle Scholar
  29. 29.
    Carlberg M, Larsson O (1993) Role of N-linked glycosylation in cell-cycle progression and initiation of DNA synthesis in tumor-transformed human fibroblasts. Anticancer Res 13:167–171PubMedGoogle Scholar
  30. 30.
    Brewer JW, Diehl JA (2000) PERK mediates cell-cycle exit during the mammalian unfolded protein response. Proc Natl Acad Sci USA 97:12625–12630PubMedCrossRefGoogle Scholar
  31. 31.
    McCullough KD, Martindale JL, Klotz LO et al (2001) Gadd153 sensitizes cells to endoplasmic reticulum stress by down-regulating Bcl2 and perturbing the cellular redox state. Mol Cell Biol 21:1249–1259PubMedCrossRefGoogle Scholar
  32. 32.
    Hou J, Wang D, Zhang R et al (2008) Experimental therapy of hepatoma with artemisinin and its derivatives: in vitro and in vivo activity, chemosensitization, and mechanisms of action. Clin Cancer Res 14:5519–5530PubMedCrossRefGoogle Scholar
  33. 33.
    Zhou HJ, Wang Z, Li A (2008) Dihydroartemisinin induces apoptosis in human leukemia cells HL60 via downregulation of transferrin receptor expression. Anticancer Drugs 19:247–255PubMedCrossRefGoogle Scholar
  34. 34.
    Wu Y, Fabritius M, Ip C (2009) Chemotherapeutic sensitization by endoplasmic reticulum stress: increasing the efficacy of taxane against prostate cancer. Cancer Biol Ther 8:146–152PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Division of Anti-tumor Pharmacology, State Key Laboratory of Drug Research, Shanghai Institute of Materia MedicaChinese Academy of SciencesShanghaiPeople’s Republic of China

Personalised recommendations