Skip to main content

Advertisement

SpringerLink
Log in

Mechanism of triptolide-induced apoptosis: effect on caspase activation and Bid cleavage and essentiality of the hydroxyl group of triptolide

  • Original Article
  • Published:
Journal of Molecular Medicine Aims and scope Submit manuscript

Abstract

Triptolide is a compound extracted from the Chinese herb Tripterygium wilfordii Hook. f. Triptolide has potent anticancer activity. However, the mechanisms by which triptolide exerts its anticancer activities remain unclear. To explore the molecular mechanisms involved in the anticancer activity of triptolide, we have examined the effect of triptolide on the growth of pancreatic carcinoma PANC-1 and cervical adenocarcinoma HeLa cells. We found that treatment of both HeLa and PANC-1 cells with triptolide potently suppressed cell growth and induced apoptosis, indicated by nuclear fragmentation and blebbing. In both HeLa and PANC-1 cells, apoptosis induced by triptolide was associated with activation of both caspase-3 and caspase-8, and cleavage of poly(ADP-ribose) polymerase and Bid. Moreover, in HeLa cells, caspase-9 is also significantly activated in response to triptolide. Overexpression of Bcl-2 in HeLa cells substantially attenuated triptolide-induced apoptosis. Interestingly, substitution of the 14-OH of triptolide with an acetyl group abrogated both its anticancer and its antiinflammatory activities. Our studies suggest that triptolide may exert its anticancer effects by initiating apoptosis through both death-receptor- and mitochondria-mediated pathways. Our results indicate that both the apoptosis-promoting and the antiinflammatory activities of triptolide depend on the 14-OH group.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

Abbreviations

PARP:

Poly(ADP-ribose) polymerase

DMSO:

Dimethyl sulfoxide

LPS:

Lipopolysaccharide

IL-1β:

Interleukin-1β

TNF-α:

Tumor necrosis factor-α

PBS:

Phosphate-buffered saline

PI:

Propidium iodide

FITC:

Fluorescein isothiocyanate

References

  1. Adams JM, Cory S (1998) The Bcl-2 protein family: arbiters of cell survival. Science 281:1322–1326

    Article  PubMed  CAS  Google Scholar 

  2. Beutler B (2000) Tlr4: central component of the sole mammalian LPS sensor. Curr Opin Immunol 12:20–26

    Article  PubMed  CAS  Google Scholar 

  3. Chan EW, Cheng SC, Sin FW, Xie Y (2001) Triptolide induced cytotoxic effects on human promyelocytic leukemia, T cell lymphoma and human hepatocellular carcinoma cell lines. Toxicol Lett 122:81–87

    Article  PubMed  CAS  Google Scholar 

  4. Chen C, Edelstein LC, Gelinas C (2000) The Rel/NF-kappaB family directly activates expression of the apoptosis inhibitor Bcl-x(L). Mol Cell Biol 20:2687–2695

    Article  PubMed  Google Scholar 

  5. Chen M, Wang J (2002) Initiator caspases in apoptosis signaling pathways. Apoptosis 7:313–319

    Article  PubMed  CAS  Google Scholar 

  6. Chen P, Li J, Barnes J, Kokkonen GC, Lee JC, Liu Y (2002) Restraint of proinflammatory cytokine biosynthesis by mitogen-activated protein kinase phosphatase-1 in lipopolysaccharide-stimulated macrophages. J Immunol 169:6408–6416

    PubMed  CAS  Google Scholar 

  7. Chen W, Martindale JL, Holbrook NJ, Liu Y (1998) Tumor promoter arsenite activates extracellular signal-regulated kinase through a signaling pathway mediated by epidermal growth factor receptor and Shc. Mol Cell Biol 18:5178–5188

    PubMed  CAS  Google Scholar 

  8. Ding GS (1987) Important Chinese herbal remedies. Clin Ther 9:345–357

    PubMed  CAS  Google Scholar 

  9. Feldmann M, Brennan FM, Foxwell BM, Maini RN (2001) The role of TNF alpha and IL-1 in rheumatoid arthritis. Curr Dir Autoimmun 3:188–199

    Article  PubMed  CAS  Google Scholar 

  10. Fidler JM, Li K, Chung C, Wei K, Ross JA, Gao M, Rosen GD (2003) PG490-88, a derivative of triptolide, causes tumor regression and sensitizes tumors to chemotherapy. Mol Cancer Ther 2:855–862

    PubMed  CAS  Google Scholar 

  11. Green DR (2005) Apoptotic pathways: ten minutes to dead. Cell 121:671–674

    Article  PubMed  CAS  Google Scholar 

  12. Han J, Thompson P, Beutler B (1990) Dexamethasone and pentoxifylline inhibit endotoxin-induced cachectin/tumor necrosis factor synthesis at separate points in the signaling pathway. J Exp Med 172:391–394

    Article  PubMed  CAS  Google Scholar 

  13. Kiviharju TM, Lecane PS, Sellers RG, Peehl DM (2002) Antiproliferative and proapoptotic activities of triptolide (PG490), a natural product entering clinical trials, on primary cultures of human prostatic epithelial cells. Clin Cancer Res 8:2666–2674

    PubMed  CAS  Google Scholar 

  14. Krishna G, Liu K, Shigemitsu H, Gao M, Raffin TA, Rosen GD (2001) PG490-88, a derivative of triptolide, blocks bleomycin-induced lung fibrosis. Am J Pathol 158:997–1004

    PubMed  CAS  Google Scholar 

  15. Kupchan SM, Court WA, Dailey RG Jr, Gilmore CJ, Bryan RF (1972) Triptolide and tripdiolide, novel antileukemic diterpenoid triepoxides from Tripterygium wilfordii. J Am Chem Soc 94:7194–7195

    Article  PubMed  CAS  Google Scholar 

  16. Lee HH, Dadgostar H, Cheng Q, Shu J, Cheng G (1999) NF-kappaB-mediated up-regulation of Bcl-x and Bfl-1/A1 is required for CD40 survival signaling in B lymphocytes. Proc Natl Acad Sci U S A 96:9136–9141

    Article  PubMed  CAS  Google Scholar 

  17. Li J, Gorospe M, Hutter D, Barnes J, Keyse SM, Liu Y (2001) Transcriptional induction of MKP-1 in response to stress is associated with histone H3 phosphorylation–acetylation. Mol Cell Biol 21:8213–8224

    Article  PubMed  CAS  Google Scholar 

  18. Li XW,Weir MR (1990) Radix Tripterygium wilfordii—a Chinese herbal medicine with potent immunosuppressive properties. Transplantation 50:82–86

    Article  PubMed  CAS  Google Scholar 

  19. Lowe SW, Schmitt EM, Smith SW, Osborne BA, Jacks T (1993) p53 is required for radiation-induced apoptosis in mouse thymocytes. Nature 362:847–849

    Article  PubMed  CAS  Google Scholar 

  20. Luo X, Budihardjo I, Zou H, Slaughter C, Wang X (1998) Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell 94:481–490

    Article  PubMed  CAS  Google Scholar 

  21. May E, Jenkins JR, May P (1991) Endogenous HeLa p53 proteins are easily detected in HeLa cells transfected with mouse deletion mutant p53 gene. Oncogene 6:1363–1365

    PubMed  CAS  Google Scholar 

  22. Mei Z, Li X, Wu Q, Hu S, Yang X (2005) The research on the anti-inflammatory activity and hepatotoxicity of triptolide-loaded solid lipid nanoparticle. Pharmacol Res 51:345–351

    Article  PubMed  CAS  Google Scholar 

  23. Qiu D, Zhao G, Aoki Y, Shi L, Uyei A, Nazarian S, Ng JC, Kao PN (1999) Immunosuppressant PG490 (triptolide) inhibits T-cell interleukin-2 expression at the level of purine-box/nuclear factor of activated T-cells and NF-kappaB transcriptional activation. J Biol Chem 274:13443–13450

    Article  PubMed  CAS  Google Scholar 

  24. Redston MS, Caldas C, Seymour AB, Hruban RH, da Costa L, Yeo CJ, Kern SE (1994) p53 mutations in pancreatic carcinoma and evidence of common involvement of homocopolymer tracts in DNA microdeletions. Cancer Res 54:3025–3033

    PubMed  CAS  Google Scholar 

  25. Reed JC (1995) Bcl-2 family proteins: regulators of chemoresistance in cancer. Toxicol Lett 82–83:155–158

    Article  PubMed  Google Scholar 

  26. Ruggeri B, Zhang SY, Caamano J, DiRado M, Flynn SD, Klein-Szanto AJ (1992) Human pancreatic carcinomas and cell lines reveal frequent and multiple alterations in the p53 and Rb-1 tumor-suppressor genes. Oncogene 7:1503–1511

    PubMed  CAS  Google Scholar 

  27. Shamon LA, Pezzuto JM, Graves JM, Mehta RR, Wangcharoentrakul S, Sangsuwan R, Chaichana S, Tuchinda P, Cleason P, Reutrakul V (1997) Evaluation of the mutagenic, cytotoxic, and antitumor potential of triptolide, a highly oxygenated diterpene isolated from Tripterygium wilfordii. Cancer Lett 112:113–117

    Article  PubMed  Google Scholar 

  28. Shepherd EG, Zhao Q, Welty SE, Hansen TN, Smith CV, Liu Y (2004) The function of mitogen-activated protein kinase phosphatase-1 in peptidoglycan-stimulated macrophages. J Biol Chem 279:54023–54031

    Article  PubMed  CAS  Google Scholar 

  29. Stehlik C, de Martin R, Kumabashiri I, Schmid JA, Binder BR, Lipp J (1998) Nuclear factor (NF)-kappaB-regulated X-chromosome-linked iap gene expression protects endothelial cells from tumor necrosis factor alpha-induced apoptosis. J Exp Med 188:211–216

    Article  PubMed  CAS  Google Scholar 

  30. Tao X, Lipsky PE (2000) The Chinese anti-inflammatory and immunosuppressive herbal remedy Tripterygium wilfordii Hook F. Rheum Dis Clin North Am 26:29–50, viii

    Article  PubMed  CAS  Google Scholar 

  31. Tao X, Cai JJ, Lipsky PE (1995) The identity of immunosuppressive components of the ethyl acetate extract and chloroform methanol extract (T2) of Tripterygium wilfordii Hook F. J Pharmacol Exp Ther 272:1305–1312

    PubMed  CAS  Google Scholar 

  32. Tao X, Cush JJ, Garret M, Lipsky PE (2001) A phase I study of ethyl acetate extract of the Chinese antirheumatic herb Tripterygium wilfordii Hook F in rheumatoid arthritis. J Rheumatol 28:2160–2167

    PubMed  CAS  Google Scholar 

  33. Tao X, Younger J, Fan FZ, Wang B, Lipsky PE (2002) Benefit of an extract of Tripterygium wilfordii Hook F in patients with rheumatoid arthritis: a double-blind, placebo-controlled study. Arthritis Rheum 46:1735–1743

    Article  PubMed  Google Scholar 

  34. Wang X, Gorospe M, Huang Y, Holbrook NJ (1997) p27Kip1 overexpression causes apoptotic death of mammalian cells. Oncogene 15:2991–2997

    Article  PubMed  CAS  Google Scholar 

  35. Wu JJ, Bennett AM (2005) Essential role for mitogen-activated protein (MAP) kinase phosphatase-1 in stress-responsive MAP kinase and cell survival signaling. J Biol Chem 280:16461–16466

    Article  PubMed  CAS  Google Scholar 

  36. Yaginuma Y, Westphal H (1991) Analysis of the p53 gene in human uterine carcinoma cell lines. Cancer Res 51:6506–6509

    PubMed  CAS  Google Scholar 

  37. Yang S, Chen J, Guo Z, Xu XM, Wang L, Pei XF, Yang J, Underhill CB, Zhang L (2003) Triptolide inhibits the growth and metastasis of solid tumors. Mol Cancer Ther 2:65–72

    PubMed  CAS  Google Scholar 

  38. Yinjun L, Jie J, Yungui W (2005) Triptolide inhibits transcription factor NF-kappaB and induces apoptosis of multiple myeloma cells. Leuk Res 29:99–105

    Article  PubMed  CAS  Google Scholar 

  39. Zhao Q, Shepherd EG, Manson ME, Nelin LD, Sorokin A, Liu Y (2005) The role of mitogen-activated protein kinase phosphatase-1 in the response of alveolar macrophages to lipopolysaccharide: attenuation of proinflammatory cytokine biosynthesis via feedback control of p38. J Biol Chem 280:8101–8108

    Article  PubMed  CAS  Google Scholar 

  40. Zong WX, Edelstein LC, Chen C, Bash J, Gelinas C (1999) The prosurvival Bcl-2 homolog Bfl-1/A1 is a direct transcriptional target of NF-kappaB that blocks TNFalpha-induced apoptosis. Genes Dev 13:382–387

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr. Lili Liu for technical assistance and Dr. Xiantao Wang for providing valuable reagents. We are grateful to Drs. John Fidler and Lurong Zhang for helpful discussion. This work was supported by a National Institute of Allergy and Infectious Diseases (NIAID) grant (R01AI57798) and by the Columbus Children's Research Institute.

Author information

Authors and Affiliations

  1. Department of Pediatrics, Center for Developmental Pharmacology and Toxicology, Children's Research Institute, The Ohio State University, Columbus, OH, 43205, USA

    Xianxi Wang, Leif D. Nelin & Yusen Liu

  2. Integrated Biomedical Science Graduate Program, The Ohio State University, Columbus, OH, 43210, USA

    Ranyia Matta & Yusen Liu

  3. Department of Chemistry, The Ohio State University, Columbus, OH, 43210, USA

    Gang Shen & Dehua Pei

Authors
  1. Xianxi Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ranyia Matta
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gang Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Leif D. Nelin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dehua Pei
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yusen Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yusen Liu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, X., Matta, R., Shen, G. et al. Mechanism of triptolide-induced apoptosis: effect on caspase activation and Bid cleavage and essentiality of the hydroxyl group of triptolide. J Mol Med 84, 405–415 (2006). https://doi.org/10.1007/s00109-005-0022-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00109-005-0022-4

Keywords

Search

Navigation