Advertisement

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

  • Xianxi Wang
  • Ranyia Matta
  • Gang Shen
  • Leif D. Nelin
  • Dehua Pei
  • Yusen LiuEmail author
Original Article

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.

Keywords

Triptolide Apoptosis Caspase Cancer Chemotherapy 

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

Notes

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.

References

  1. 1.
    Adams JM, Cory S (1998) The Bcl-2 protein family: arbiters of cell survival. Science 281:1322–1326PubMedCrossRefGoogle Scholar
  2. 2.
    Beutler B (2000) Tlr4: central component of the sole mammalian LPS sensor. Curr Opin Immunol 12:20–26PubMedCrossRefGoogle Scholar
  3. 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 122:81–87PubMedCrossRefGoogle Scholar
  4. 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 20:2687–2695PubMedCrossRefGoogle Scholar
  5. 5.
    Chen M, Wang J (2002) Initiator caspases in apoptosis signaling pathways. Apoptosis 7:313–319PubMedCrossRefGoogle Scholar
  6. 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 169:6408–6416PubMedGoogle Scholar
  7. 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 18:5178–5188PubMedGoogle Scholar
  8. 8.
    Ding GS (1987) Important Chinese herbal remedies. Clin Ther 9:345–357PubMedGoogle Scholar
  9. 9.
    Feldmann M, Brennan FM, Foxwell BM, Maini RN (2001) The role of TNF alpha and IL-1 in rheumatoid arthritis. Curr Dir Autoimmun 3:188–199PubMedCrossRefGoogle Scholar
  10. 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 2:855–862PubMedGoogle Scholar
  11. 11.
    Green DR (2005) Apoptotic pathways: ten minutes to dead. Cell 121:671–674PubMedCrossRefGoogle Scholar
  12. 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 172:391–394PubMedCrossRefGoogle Scholar
  13. 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 8:2666–2674PubMedGoogle Scholar
  14. 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 158:997–1004PubMedGoogle Scholar
  15. 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 94:7194–7195PubMedCrossRefGoogle Scholar
  16. 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 96:9136–9141PubMedCrossRefGoogle Scholar
  17. 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 21:8213–8224PubMedCrossRefGoogle Scholar
  18. 18.
    Li XW,Weir MR (1990) Radix Tripterygium wilfordii—a Chinese herbal medicine with potent immunosuppressive properties. Transplantation 50:82–86PubMedCrossRefGoogle Scholar
  19. 19.
    Lowe SW, Schmitt EM, Smith SW, Osborne BA, Jacks T (1993) p53 is required for radiation-induced apoptosis in mouse thymocytes. Nature 362:847–849PubMedCrossRefGoogle Scholar
  20. 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 94:481–490PubMedCrossRefGoogle Scholar
  21. 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 6:1363–1365PubMedGoogle Scholar
  22. 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 51:345–351PubMedCrossRefGoogle Scholar
  23. 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 274:13443–13450PubMedCrossRefGoogle Scholar
  24. 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 54:3025–3033PubMedGoogle Scholar
  25. 25.
    Reed JC (1995) Bcl-2 family proteins: regulators of chemoresistance in cancer. Toxicol Lett 82–83:155–158PubMedCrossRefGoogle Scholar
  26. 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 7:1503–1511PubMedGoogle Scholar
  27. 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 112:113–117PubMedCrossRefGoogle Scholar
  28. 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 279:54023–54031PubMedCrossRefGoogle Scholar
  29. 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 188:211–216PubMedCrossRefGoogle Scholar
  30. 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, viiiPubMedCrossRefGoogle Scholar
  31. 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 272:1305–1312PubMedGoogle Scholar
  32. 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 28:2160–2167PubMedGoogle Scholar
  33. 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 46:1735–1743PubMedCrossRefGoogle Scholar
  34. 34.
    Wang X, Gorospe M, Huang Y, Holbrook NJ (1997) p27Kip1 overexpression causes apoptotic death of mammalian cells. Oncogene 15:2991–2997PubMedCrossRefGoogle Scholar
  35. 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 280:16461–16466PubMedCrossRefGoogle Scholar
  36. 36.
    Yaginuma Y, Westphal H (1991) Analysis of the p53 gene in human uterine carcinoma cell lines. Cancer Res 51:6506–6509PubMedGoogle Scholar
  37. 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 2:65–72PubMedGoogle Scholar
  38. 38.
    Yinjun L, Jie J, Yungui W (2005) Triptolide inhibits transcription factor NF-kappaB and induces apoptosis of multiple myeloma cells. Leuk Res 29:99–105PubMedCrossRefGoogle Scholar
  39. 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 280:8101–8108PubMedCrossRefGoogle Scholar
  40. 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 13:382–387PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Xianxi Wang
    • 1
  • Ranyia Matta
    • 2
  • Gang Shen
    • 3
  • Leif D. Nelin
    • 1
  • Dehua Pei
    • 3
  • Yusen Liu
    • 1
    • 2
    Email author
  1. 1.Department of Pediatrics, Center for Developmental Pharmacology and Toxicology, Children's Research InstituteThe Ohio State UniversityColumbusUSA
  2. 2.Integrated Biomedical Science Graduate ProgramThe Ohio State UniversityColumbusUSA
  3. 3.Department of ChemistryThe Ohio State UniversityColumbusUSA

Personalised recommendations