Skip to main content

Advertisement

SpringerLink
Log in

Gambogic acid is cytotoxic to cancer cells through inhibition of the ubiquitin-proteasome system

  • PRECLINICAL STUDIES
  • Published:
Investigational New Drugs Aims and scope Submit manuscript

Summary

Gambogic acid (GA), displays cytotoxicity towards a wide variety of tumor cells and has been shown to affect many important cell-signaling pathways. In the present work, we investigated the mechanism of action of GA by analysis of drug-induced changes in gene expression profiles and identified GA and the derivative dihydro GA as possible inhibitors of the ubiquitin-proteasome system (UPS). Both GA and dihydro GA inhibited proteasome function in cells resulting in the accumulation of polyubiquitin complexes. In vitro experiments showed that both GA and dihydro GA inhibited 20S chymotrypsin activity and the inhibitory effects of GA and dihydro GA on proteasome function corresponded with apoptosis induction and cell death. In conclusion, our results show that GA and dihydro GA exert their cytotoxic activity through inhibition of the UPS, specifically by acting as inhibitors of the chymotrypsin activity of the 20S proteasome.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Similar content being viewed by others

References

  1. Cragg GM, Grothaus PG, Newman DJ (2009) Impact of natural products on developing new anti-cancer agents. Chem Rev 109:3012–3043

    Article  PubMed  CAS  Google Scholar 

  2. Chen J, Gu HY, Lu N, Yang Y, Liu W, Qi Q, Rong JJ, Wang XT, You QD, Guo QL (2008) Microtubule depolymerization and phosphorylation of c-Jun N-terminal kinase-1 and p38 were involved in gambogic acid induced cell cycle arrest and apoptosis in human breast carcinoma MCF-7 cells. Life Sci 83:103–109

    Article  PubMed  CAS  Google Scholar 

  3. Zhang HZ, Kasibhatla S, Wang Y, Herich J, Guastella J, Tseng B, Drewe J, Cai SX (2004) Discovery, characterization and SAR of gambogic acid as a potent apoptosis inducer by a HTS assay. Bioorg Med Chem 12:309–317

    Article  PubMed  Google Scholar 

  4. Reutrakul V, Anantachoke N, Pohmakotr M, Jaipetch T, Sophasan S, Yoosook C, Kasisit J, Napaswat C, Santisuk T, Tuchinda P (2007) Cytotoxic and anti-HIV-1 caged xanthones from the resin and fruits of Garcinia hanburyi. Planta Med 73:33–40

    Article  PubMed  CAS  Google Scholar 

  5. Wu ZQ, Guo QL, You QD, Zhao L, Gu HY (2004) Gambogic acid inhibits proliferation of human lung carcinoma SPC-A1 cells in vivo and in vitro and represses telomerase activity and telomerase reverse transcriptase mRNA expression in the cells. Biol Pharm Bull 27:1769–1774

    Article  PubMed  CAS  Google Scholar 

  6. Yi T, Yi Z, Cho SG, Luo J, Pandey MK, Aggarwal BB, Liu M (2008) Gambogic acid inhibits angiogenesis and prostate tumor growth by suppressing vascular endothelial growth factor receptor 2 signaling. Cancer Res 68:1843–1850

    Article  PubMed  CAS  Google Scholar 

  7. Qi Q, Gu H, Yang Y, Lu N, Zhao J, Liu W, Ling H, You QD, Wang X, Guo Q (2008) Involvement of matrix metalloproteinase 2 and 9 in gambogic acid induced suppression of MDA-MB-435 human breast carcinoma cell lung metastasis. J Mol Med 86:1367–1377

    Article  PubMed  CAS  Google Scholar 

  8. Qin Y, Meng L, Hu C, Duan W, Zuo Z, Lin L, Zhang X, Ding J (2007) Gambogic acid inhibits the catalytic activity of human topoisomerase IIalpha by binding to its ATPase domain. Mol Cancer Ther 6:2429–2440

    Article  PubMed  CAS  Google Scholar 

  9. Pandey MK, Sung B, Ahn KS, Kunnumakkara AB, Chaturvedi MM, Aggarwal BB (2007) Gambogic acid, a novel ligand for transferrin receptor, potentiates TNF-induced apoptosis through modulation of the nuclear factor-kappaB signaling pathway. Blood 110:3517–3525

    Article  PubMed  CAS  Google Scholar 

  10. Zhai D, Jin C, Shiau CW, Kitada S, Satterthwait AC, Reed JC (2008) Gambogic acid is an antagonist of antiapoptotic Bcl-2 family proteins. Mol Cancer Ther 7:1639–1646

    Article  PubMed  CAS  Google Scholar 

  11. Cvek B, Dvorak Z (2011) The ubiquitin-proteasome system (UPS) and the mechanism of action of bortezomib. Curr Pharm Des 17:1483–1499

    Article  PubMed  CAS  Google Scholar 

  12. Tsukamoto S, Yokosawa H (2009) Targeting the proteasome pathway. Expert Opin Ther Targets 13:605–621

    Article  PubMed  CAS  Google Scholar 

  13. Adams J (2004) The proteasome: a suitable antineoplastic target. Nat Rev Cancer 4:349–360

    Article  PubMed  CAS  Google Scholar 

  14. Peters JM, Cejka Z, Harris JR, Kleinschmidt JA, Baumeister W (1993) Structural features of the 26 S proteasome complex. J Mol Biol 234:932–937

    Article  PubMed  CAS  Google Scholar 

  15. da Fonseca PC, He J, Morris EP (2012) Molecular model of the human 26S proteasome. Mol Cell 46:54–66

    Article  PubMed  Google Scholar 

  16. An B, Goldfarb RH, Siman R, Dou QP (1998) Novel dipeptidyl proteasome inhibitors overceome Bcl-2 protective function and selectively acculmulate the cyclin-dependent kinase inhibitor p27 and induce apoptosis in transformed, but not normal, human fibroblasts. Cell Death Differ 5:1062–1975

    Article  PubMed  CAS  Google Scholar 

  17. Hideshima T, Richardson P, Chauhan D, Palombella VJ, Elliott PJ, Adams J, Anderson KC (2001) The protesaome inhibitor PS-341 inhibits growth, induces apoptosis, and overcomes drug resitance in human multiple myeloma cells. Cancer Res. 61

  18. Tsukamoto S, Yokosawa H (2010) Inhibition of the ubiquitin-proteasome system by natural products for cancer therapy. Planta Med 76:1064–1074

    Article  PubMed  CAS  Google Scholar 

  19. Potts BC, Albitar MX, Anderson KC, Baritaki S, Berkers C, Bonavida B, Chandra J, Chauhan D, Cusack JC, Fenical W, Ghobiral IM, Groll M, Jensen PR, Lam KS, Lloyd GK, McBride W, McConkey DJ, Miller CP, Neuteboom STC, Oki Y, Ovaa H, Pajonk F, Richardson PG, Roccaro AM, Sloss CM, Spear MA, Valashi E, Younes A, Palladino MA (2011) Marizomib, a proteasome inhibitor for all seasons: preclinical profile and a framework for clinical trials. Curr Cancer Drug Targets 11:254–284

    Article  CAS  Google Scholar 

  20. Kuhn DJ, Orlowski RZ, Bjorklund CC (2011) Second generation proteasome inhibitors: carfilzomib and Immunoproteasome-Specific Inhibitors (IPSIs). Curr Cancer Drug Targets 11:285–295

    Article  PubMed  CAS  Google Scholar 

  21. Yang H, Landis-Piwowar K, Chan TH, Dou QP (2011) Green tea polyphenols as proteasome inhibitors: implication in chemoprevention. Curr Cancer Drug Targets 11:296–306

    Article  PubMed  CAS  Google Scholar 

  22. Cecarini V, Cuccioloni M, Mozzicafreddo M, Bonfili L, Angeletti M, Eleuteri AM (2011) Targeting proteasome with naturally occurring compounds in cancer treatment. Curr Cancer Drug Targets 11:307–324

    Article  PubMed  CAS  Google Scholar 

  23. Rickardson L, Wickstrom M, Larsson R, Lovborg H (2007) Image-based screening for the identification of novel proteasome inhibitors. J Biomol Screen 12:203–210

    Article  PubMed  CAS  Google Scholar 

  24. Lavelin I, Beer A, Kam Z, Rotter V, Oren M, Navon A, Geiger B (2009) Discovery of novel proteasome inhibitors using a high-content cell-based screening system. PLoS One 4:e8503

    Article  PubMed  Google Scholar 

  25. Menendez-Benito V, Verhoef LG, Masucci MG, Dantuma NP (2005) Endoplasmic reticulum stress compromises the ubiquitin-proteasome system. Hum Mol Genet 14:2787–2799

    Article  PubMed  CAS  Google Scholar 

  26. Dhar S, Nygren P, Csoka K, Botling J, Nilsson K, Larsson R (1996) Anti-cancer drug characterisation using a human cell line panel representing defined types of drug resistance. Br J Cancer 74:888–896

    Article  PubMed  CAS  Google Scholar 

  27. Rickardson L, Fryknas M, Dhar S, Lovborg H, Gullbo J, Rydaker M, Nygren P, Gustafsson MG, Larsson R, Isaksson A (2005) Identification of molecular mechanisms for cellular drug resistance by combining drug activity and gene expression profiles. Br J Cancer 93:483–492

    Article  PubMed  CAS  Google Scholar 

  28. Lamb J, Crawford ED, Peck D, Modell JW, Blat IC, Wrobel MJ, Lerner J, Brunet JP, Subramanian A, Ross KN, Reich M, Hieronymus H, Wei G, Armstrong SA, Haggarty SJ, Clemons PA, Wei R, Carr SA, Lander ES, Golub TR (2006) The Connectivity Map: using gene-expression signatures to connect small molecules, genes, and disease. Science 313:1929–1935

    Article  PubMed  CAS  Google Scholar 

  29. Fayad W, Fryknas M, Brnjic S, Olofsson MH, Larsson R, Linder S (2009) Identification of a novel topoisomerase inhibitor effective in cells overexpressing drug efflux transporters. PLoS One 4:e7238

    Article  PubMed  Google Scholar 

  30. Larsson R, Nygren P (1989) A rapid fluorometric method for semiautomated determination of cytotoxicity and cellular proliferation of human tumor cell lines in microculture. Anticancer Res 9:1111–1119

    PubMed  CAS  Google Scholar 

  31. Lindhagen E, Nygren P, Larsson R (2008) The fluorometric microculture cytotoxicity assay. Nat Protoc 3:1364–1369

    Article  PubMed  CAS  Google Scholar 

  32. Paull KD, Shoemaker RH, Hodes L, Monks A, Scudiero DA, Rubinstein L, Plowman J, Boyd MR (1989) Display and analysis of patterns of differential activity of drugs against human tumor cell lines: development of mean graph and COMPARE algorithm. J Natl Cancer Inst 81:1088–1092

    Article  PubMed  CAS  Google Scholar 

  33. Shibata T, Yamada T, Kondo M, Tanahashi N, Tanaka K, Nakamura H, Masutani H, Yodoi J, Uchida K (2003) An endogenous electrophile that modulates the regulatory mechanism of protein turnover: inhibitory effects of 15-deoxy-Delta 12,14-prostaglandin J2 on proteasome. Biochemistry 42:13960–13968

    Article  PubMed  CAS  Google Scholar 

  34. Yang H, Chen D, Cui QC, Yuan X, Dou QP (2006) Celastrol, a triterpene extracted from the Chinese "Thunder of God Vine," is a potent proteasome inhibitor and suppresses human prostate cancer growth in nude mice. Cancer Res 66:4758–4765

    Article  PubMed  CAS  Google Scholar 

  35. Yang H, Shi G, Dou QP (2007) The tumor proteasome is a primary target for the natural anticancer compound Withaferin A isolated from "Indian winter cherry". Mol Pharmacol 71:426–437

    Article  PubMed  CAS  Google Scholar 

  36. Lovborg H, Oberg F, Rickardson L, Gullbo J, Nygren P, Larsson R (2006) Inhibition of proteasome activity, nuclear factor-KappaB translocation and cell survival by the antialcoholism drug disulfiram. Int J Cancer 118:1577–1580

    Article  PubMed  Google Scholar 

  37. Zimmermann J, Erdmann D, Lalande I, Grossenbacher R, Noorani M, Furst P (2000) Proteasome inhibitor induced gene expression profiles reveal overexpression of transcriptional regulators ATF3, GADD153 and MAD1. Oncogene 19:2913–2920

    Article  PubMed  CAS  Google Scholar 

  38. Yew EH, Cheung NS, Choy MS, Qi RZ, Lee AY, Peng ZF, Melendez AJ, Manikandan J, Koay ES, Chiu LL, Ng WL, Whiteman M, Kandiah J, Halliwell B (2005) Proteasome inhibition by lactacystin in primary neuronal cells induces both potentially neuroprotective and pro-apoptotic transcriptional responses: a microarray analysis. J Neurochem 94:943–956

    Article  PubMed  CAS  Google Scholar 

  39. Mitsiades N, Mitsiades CS, Poulaki V, Chauhan D, Fanourakis G, Gu X, Bailey C, Joseph M, Libermann TA, Treon SP, Munshi NC, Richardson PG, Hideshima T, Anderson KC (2002) Molecular sequelae of proteasome inhibition in human multiple myeloma cells. Proc Natl Acad Sci USA 99:14374–14379

    Article  PubMed  CAS  Google Scholar 

  40. Fribley A, Zeng Q, Wang CY (2004) Proteasome inhibitor PS-341 induces apoptosis through induction of endoplasmic reticulum stress-reactive oxygen species in head and neck squamous cell carcinoma cells. Mol Cell Biol 24:9695–9704

    Article  PubMed  CAS  Google Scholar 

  41. Hong SH, Kim J, Kim JM, Lee SY, Shin DS, Son KH, Han DC, Sung YK, Kwon BM (2007) Apoptosis induction of 2'-hydroxycinnamaldehyde as a proteasome inhibitor is associated with ER stress and mitochondrial perturbation in cancer cells. Biochem Pharmacol 74:557–565

    Article  PubMed  CAS  Google Scholar 

  42. Noonan EJ, Place RF, Giardina C, Hightower LE (2007) Hsp70B' regulation and function. Cell Stress Chaperones 12:393–402

    Article  PubMed  Google Scholar 

  43. Wu WT, Chi KH, Ho FM, Tsao WC, Lin WW (2004) Proteasome inhibitors up-regulate haem oxygenase-1 gene expression: requirement of p38 MAPK (mitogen-activated protein kinase) activation but not of NF-kappaB (nuclear factor kappaB) inhibition. Biochem J 379:587–593

    Article  PubMed  CAS  Google Scholar 

  44. Shen J, Reis J, Morrison DC, Papasian C, Raghavakaimal S, Kolbert C, Qureshi AA, Vogel SN, Qureshi N (2006) Key inflammatory signaling pathways are regulated by the proteasome. Shock 25:472–484

    Article  PubMed  CAS  Google Scholar 

  45. Tang ZY, Wu YL, Gao SL, Shen HW (2008) Effects of the proteasome inhibitor bortezomib on gene expression profiles of pancreatic cancer cells. J Surg Res 145:111–123

    Article  PubMed  Google Scholar 

  46. Kinyamu HK, Collins JB, Grissom SF, Hebbar PB, Archer TK (2008) Genome wide transcriptional profiling in breast cancer cells reveals distinct changes in hormone receptor target genes and chromatin modifying enzymes after proteasome inhibition. Mol Carcinog 47:845–885

    Article  PubMed  CAS  Google Scholar 

  47. Kasibhatla S, Jessen KA, Maliartchouk S, Wang JY, English NM, Drewe J, Qiu L, Archer SP, Ponce AE, Sirisoma N, Jiang S, Zhang HZ, Gehlsen KR, Cai SX, Green DR, Tseng B (2005) A role for transferrin receptor in triggering apoptosis when targeted with gambogic acid. Proc Natl Acad Sci USA 102:12095–12100

    Article  PubMed  CAS  Google Scholar 

  48. Kapuria V, Peterson LF, Fang D, Bornmann WG, Talpaz M, Donato NJ (2010) Deubiquitinase inhibition by small-molecule WP1130 triggers aggresome formation and tumor cell apoptosis. Cancer Res 70:9265–9276

    Article  PubMed  CAS  Google Scholar 

  49. Koulich E, Li X, DeMartino GN (2008) Relative structural and functional roles of multiple deubiquitylating proteins associated with mammalian 26S proteasome. Mol Biol Cell 19:1072–1082

    Article  PubMed  CAS  Google Scholar 

  50. D'Arcy P, Brnjic S, Olofsson MH, Fryknas M, Lindsten K, De Cesare M, Perego P, Sadeghi B, Hassan M, Larsson R, Linder S (2011) Inhibition of proteasome deubiquitinating activity as a new cancer therapy. Nat Med 17:1630–1640

    Article  Google Scholar 

  51. Batova A, Altomare D, Chantarasriwong O, Ohlsen KL, Creek KE, Lin YC, Messersmith A, Yu AL, Yu J, Theodorakis EA (2010) The synthetic caged garcinia xanthone cluvenone induces cell stress and apoptosis and has immune modulatory activity. Mol Cancer Ther 9:2869–2878

    Article  PubMed  CAS  Google Scholar 

  52. Guo QL, Lin SS, You QD, Gu HY, Yu J, Zhao L, Qi Q, Liang F, Tan Z, Wang X (2006) Inhibition of human telomerase reverse transcriptase gene expression by gambogic acid in human hepatoma SMMC-7721 cells. Life Sci 78:1238–1245

    Article  PubMed  CAS  Google Scholar 

  53. Zhang L, Yi Y, Chen J, Sun Y, Guo Q, Zheng Z, Song S (2010) Gambogic acid inhibits Hsp90 and deregulates TNF-alpha/NF-kappaB in HeLa cells. Biochem Biophys Res Commun 403:282–287

    Article  PubMed  CAS  Google Scholar 

  54. Davenport J, Manjarrez JR, Peterson L, Krumm B, Blagg BS, Matts RL (2011) Gambogic acid, a natural product inhibitor of Hsp90. J Nat Prod 27:1085–1092

    Article  Google Scholar 

  55. Sethi G, Ahn KS, Pandey MK, Aggarwal BB (2007) Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-kappaB-regulated gene products and TAK1-mediated NF-kappaB activation. Blood 109:2727–2735

    PubMed  CAS  Google Scholar 

  56. Sartore-Bianchi A, Gasparri F, Galvani A, Nici L, Darnowski JW, Barbone D, Fennell DA, Gaudino G, Porta C, Mutti L (2007) Bortezomib inhibits nuclear factor-kappaB dependent survival and has potent in vivo activity in mesothelioma. Clin Cancer Res 13:5942–5951

    Article  PubMed  CAS  Google Scholar 

  57. Broemer M, Krappmann D, Scheidereit C (2004) Requirement of Hsp90 activity for IkappaB kinase (IKK) biosynthesis and for constitutive and inducible IKK and NF-kappaB activation. Oncogene 23:5378–5386

    Article  PubMed  CAS  Google Scholar 

  58. Sugita T, Tanaka S, Murakami T, Miyoshi H, Ohnuki T (1999) Immunosupressive effects of the heat shock protein 90-binding antibiotic geldanamycin. Biochem Mol Biol Int 47:587–595

    PubMed  CAS  Google Scholar 

  59. Huang H, Chen D, Li S, Li X, Liu N, Lu X, Liu S, Zhao K, Zhao C, Guo H, Yang C, Zhou P, Dong X, Zhang C, Guanmei, Dou QP, Liu J (2011) Gambogic acid enhances proteasome inhibitor-induced anticancer activity. Cancer Lett 301:221–228

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors are very grateful to Professor Nico Dantuma for the kind gift of MelJuSo UbG76V-YFP cells. We also wish to thank Maria Rydåker, Anna Haukkula and Hanna Göransson Kultima at the Uppsala Array Platform for microarray analysis, Sandra Erlund for assistance in the literature survey, and Lena Lenhammar for RNA preparation. This work was supported by unconditional grants from the Lions Cancer Research Fund and a Swedish Institute scholarship for Karolina Lesiak-Mieczkowska.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Division of Pharmacognosy, Department of Medicinal Chemistry, Biomedical Center, Uppsala University, Uppsala, Sweden

    Jenny Felth & Lars Bohlin

  2. Cancer Center Karolinska, Department of Oncology-Pathology, Karolinska Institute, Stockholm, Sweden

    Karolina Lesiak-Mieczkowska, Padraig D’Arcy & Stig Linder

  3. Division of Clinical Pharmacology, Department of Medical Sciences, Uppsala University, Uppsala, 751 85, Sweden

    Caroline Haglund, Joachim Gullbo, Rolf Larsson, Mårten Fryknäs & Linda Rickardson

Authors
  1. Jenny Felth
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Karolina Lesiak-Mieczkowska
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Padraig D’Arcy
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Caroline Haglund
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Joachim Gullbo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Rolf Larsson
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Stig Linder
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Lars Bohlin
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Mårten Fryknäs
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Linda Rickardson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Linda Rickardson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Felth, J., Lesiak-Mieczkowska, K., D’Arcy, P. et al. Gambogic acid is cytotoxic to cancer cells through inhibition of the ubiquitin-proteasome system. Invest New Drugs 31, 587–598 (2013). https://doi.org/10.1007/s10637-012-9902-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10637-012-9902-y

Keywords

Search

Navigation