Skip to main content
SpringerLink
Log in

Carnosic acid modulates Akt/IKK/NF-κB signaling by PP2A and induces intrinsic and extrinsic pathway mediated apoptosis in human prostate carcinoma PC-3 cells

  • Original Paper
  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

This study investigates the efficacy of carnosic acid (CA), a polyphenolic diterpene, isolated from the plant rosemary (Rosemarinus officinalis), on androgen-independent human prostate cancer PC-3 cells. CA induced anti-proliferative effects in PC-3 cells in a concentration- and time-dependent manner, which was due to apoptotic induction as evident from flow-cytometry, DNA laddering and TUNEL assay. Apoptosis was associated with the activation of caspase-8, -9, -3 and -7, increase in Bax:Bcl-2 ratio, release of cytochrome-c and decrease in expression of inhibitor of apoptosis (IAP) family of proteins. Apoptosis was attenuated upon pretreatment with specific inhibitors of caspase-8 (Z-IETD-fmk) and caspase-9 (Z-LEHD-fmk) suggesting the involvement of both intrinsic and extrinsic apoptotic cascades. Further, apoptosis resulted from the inhibition of IKK/NF-κB pathway as evident from decreased DNA binding activity, nuclear translocation of p50 and p65 and IκBα phosphorylation. The down-regulation of IKK/NF-κB was associated with inhibition of Akt phosphorylation and its kinase activity with a concomitant increase in the serine/threonine protein phosphatase 2A (PP2A) activity. Pharmacologic inhibition of PP2A by okadaic acid and calyculin A, significantly reversed CA-mediated apoptotic events in PC-3 cells indicating that CA induced apoptosis by activation of PP2A through modulation of Akt/IKK/NF-κB pathway. In addition, CA induced apoptosis in another androgen refractory prostate cancer DU145 cells via intrinsic pathway as evidenced from the activation of caspase 3, cleavage of PARP, increase in Bax:Bcl-2 ratio and cytochrome-c release. Carnosic acid, therefore, may have the potential for use in the prevention and/or treatment of prostate cancer.

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

Similar content being viewed by others

Abbreviations

CA:

Carnosic acid

PARP:

Poly (ADP)-ribose polymerase

IAP:

Inhibitor of apoptosis

IKK:

IκB kinase

MTT:

(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NF-κB:

Nuclear factor-κB

PI3-K:

Phosphoinositide 3-kinase

PP2A:

Protein phosphatase 2A

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling

References

  1. Jemal A, Siegel R, Xu J, Ward E (2010) Cancer statistics, 2010. CA Cancer J Clin 60:277–300

    Article  PubMed  Google Scholar 

  2. Cooperberg MR, Grossfeld GD, Lubeck DP, Carroll PR (2003) National practice patterns and time trends in androgen ablation for localized prostate cancer. J Natl Cancer Inst 95:981–989

    Article  PubMed  Google Scholar 

  3. Feldman BJ, Feldman D (2001) The development of androgen-independent prostate cancer. Nat Rev Cancer 1:34–45

    Article  PubMed  CAS  Google Scholar 

  4. Hsing AW, Devesa SS (2001) Trends and patterns of prostate cancer: what do they suggest? Epidemiol Rev 23:3–13

    Article  PubMed  CAS  Google Scholar 

  5. Syed DN, Khan N, Afaq F, Mukhtar H (2007) Chemoprevention of prostate cancer through dietary agents: progress and promise. Cancer Epidemiol Biomarkers Prev 16:2193–2203

    Article  PubMed  CAS  Google Scholar 

  6. Khan N, Afaq F, Mukhtar H (2008) Cancer chemoprevention through dietary antioxidants: progress and promise. Antioxid Redox Signal 10:475–510

    Article  PubMed  CAS  Google Scholar 

  7. Singletary KW, Nelshoppen JM (1991) Inhibition of 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumorigenesis and of in vivo formation of mammary DMBA-DNA adducts by rosemary extract. Cancer Lett 60:169–175

    Article  PubMed  CAS  Google Scholar 

  8. Singletary KW, Rokusek JT (1997) Tissue-specific enhancement of xenobiotic detoxification enzymes in mice by dietary rosemary extract. Plant Foods Hum Nutr 50:47–53

    Article  PubMed  CAS  Google Scholar 

  9. Pesakhov S, Khanin M, Studzinski GP, Danilenko M (2010) Distinct combinatorial effects of the plant polyphenols curcumin, carnosic acid, and silibinin on proliferation and apoptosis in acute myeloid leukemia cells. Nutr Cancer 62:811–824

    Article  PubMed  CAS  Google Scholar 

  10. Yu YM, Lin CH, Chan HC, Tsai HD (2009) Carnosic acid reduces cytokine-induced adhesion molecules expression and monocyte adhesion to endothelial cells. Eur J Nutr 48:101–106

    Article  PubMed  CAS  Google Scholar 

  11. Hengartner MO (2000) The biochemistry of apoptosis. Nature 407:770–776

    Article  PubMed  CAS  Google Scholar 

  12. 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 

  13. Delivani P, Martin SJ (2006) Mitochondrial membrane remodeling in apoptosis: an inside story. Cell Death Differ 13:2007–2010

    Article  PubMed  CAS  Google Scholar 

  14. Vivanco I, Sawyers CL (2002) The phosphatidylinositol 3-Kinase AKT pathway in human cancer. Nat Rev Cancer 2:489–501

    Article  PubMed  CAS  Google Scholar 

  15. Aggarwal BB (2004) Nuclear factor-kappaB: the enemy within. Cancer Cell 6:203–208

    Article  PubMed  CAS  Google Scholar 

  16. Pahl HL (1999) Activators and target genes of Rel/NF-kappaB transcription factors. Oncogene 18:6853–6866

    Article  PubMed  CAS  Google Scholar 

  17. Sovak MA, Arsura M, Zanieski G, Kavanagh KT, Sonenshein GE (1999) The inhibitory effects of transforming growth factor beta1 on breast cancer cell proliferation are mediated through regulation of aberrant nuclear factor-kappaB/Rel expression. Cell Growth Differ 10:537–544

    PubMed  CAS  Google Scholar 

  18. Lind DS, Hochwald SN, Malaty J et al (2001) Nuclear factor-kappa B is upregulated in colorectal cancer. Surgery 130:363–369

    Article  PubMed  CAS  Google Scholar 

  19. Koumakpayi IH, Le Page C, Mes-Masson AM, Saad F (2010) Hierarchical clustering of immunohistochemical analysis of the activated ErbB/PI3K/Akt/NF-kappaB signalling pathway and prognostic significance in prostate cancer. Br J Cancer 102:1163–1173

    Article  PubMed  CAS  Google Scholar 

  20. Le Page C, Koumakpayi IH, Lessard L, Saad F, Mes-Masson AM (2005) Independent role of phosphoinositol-3-kinase (PI3K) and casein kinase II (CK-2) in EGFR and Her-2-mediated constitutive NF-kappaB activation in prostate cancer cells. Prostate 65:306–315

    Article  PubMed  Google Scholar 

  21. Cohen PT (1997) Novel protein serine/threonine phosphatases: variety is the spice of life. Trends Biochem Sci 22:245–251

    Article  PubMed  CAS  Google Scholar 

  22. Van Kanegan MJ, Adams DG, Wadzinski BE, Strack S (2005) Distinct protein phosphatase 2A heterotrimers modulate growth factor signaling to extracellular signal-regulated kinases and Akt. J Biol Chem 280:36029–36036

    Article  PubMed  Google Scholar 

  23. Yu S, Shen G, Khor TO, Kim JH, Kong AN (2008) Curcumin inhibits Akt/mammalian target of rapamycin signaling through protein phosphatase-dependent mechanism. Mol Cancer Ther 7:2609–2620

    Article  PubMed  CAS  Google Scholar 

  24. Matsuoka Y, Nagahara Y, Ikekita M, Shinomiya T (2003) A novel immunosuppressive agent FTY720 induced Akt dephosphorylation in leukemia cells. Br J Pharmacol 138:1303–1312

    Article  PubMed  CAS  Google Scholar 

  25. Yesil-Celiktas O, Sevimli C, Bedir E, Vardar-Sukan F (2010) Inhibitory effects of rosemary extracts, carnosic acid and rosmarinic acid on the growth of various human cancer cell lines. Plant Foods Hum Nutr 65:158–163

    Article  PubMed  CAS  Google Scholar 

  26. Ukil A, Biswas A, Das T, Das PK (2005) 18 Beta-glycyrrhetinic acid triggers curative Th1 response and nitric oxide up-regulation in experimental visceral leishmaniasis associated with the activation of NF-kappa B. J Immunol 175:1161–1169

    PubMed  CAS  Google Scholar 

  27. Deveraux QL, Reed JC (1999) IAP family proteins—suppressors of apoptosis. Genes Dev 13:239–252

    Article  PubMed  CAS  Google Scholar 

  28. Chendil D, Das A, Dey S, Mohiuddin M, Ahmed MM (2002) Par-4, a pro-apoptotic gene, inhibits radiation-induced NF kappa B activity and Bcl-2 expression leading to induction of radiosensitivity in human prostate cancer cells PC-3. Cancer Biol Ther 1:152–160

    PubMed  CAS  Google Scholar 

  29. Vermeulen L, Vanden Berghe W, Haegeman G (2006) Regulation of NF-kappaB transcriptional activity. Cancer Treat Res 130:89–102

    Article  PubMed  CAS  Google Scholar 

  30. Vlietstra RJ, van Alewijk DC, Hermans KG, van Steenbrugge GJ, Trapman J (1998) Frequent inactivation of PTEN in prostate cancer cell lines and xenografts. Cancer Res 58:2720–2723

    PubMed  CAS  Google Scholar 

  31. Fulda S, Debatin KM (2006) Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene 25:4798–4811

    Article  PubMed  CAS  Google Scholar 

  32. Hyer ML, Shi R, Krajewska M et al (2008) Apoptotic activity and mechanism of 2-cyano-3,12-dioxoolean-1,9-dien-28-oic-acid and related synthetic triterpenoids in prostate cancer. Cancer Res 68:2927–2933

    Article  PubMed  CAS  Google Scholar 

  33. Baron A, Mancini M, Caldwell E, Cabrelle A, Bernardi P, Pagano F (2009) Serenoa repens extract targets mitochondria and activates the intrinsic apoptotic pathway in human prostate cancer cells. BJU Int 103:1275–1283

    Article  PubMed  Google Scholar 

  34. Singh AV, Xiao D, Lew KL, Dhir R, Singh SV (2004) Sulforaphane induces caspasemediated apoptosis in cultured PC-3 human prostate cancer cells and retards growth of PC-3 xenografts in vivo. Carcinogenesis 25:83–90

    Article  PubMed  CAS  Google Scholar 

  35. Gasparian AV, Yao YJ, Kowalczyk D et al (2002) The role of IKK in constitutive activation of NF-kappaB transcription factor in prostate carcinoma cells. J Cell Sci 115:141–151

    PubMed  CAS  Google Scholar 

  36. Ross JS, Jennings TA, Nazeer T et al (2003) Prognostic factors in prostate cancer. Am J Clin Pathol 120(Suppl):S85–S100

    PubMed  Google Scholar 

  37. Fradet V, Lessard L, Begin LR, Karakiewicz P, Masson AM, Saad F (2004) Nuclear factor-kappaB nuclear localization is predictive of biochemical recurrence in patients with positive margin prostate cancer. Clin Cancer Res 10:8460–8464

    Article  PubMed  CAS  Google Scholar 

  38. Suh J, Payvandi F, Edelstein LC et al (2002) Mechanisms of constitutive NF-kappaB activation in human prostate cancer cells. Prostate 52:183–200

    Article  PubMed  CAS  Google Scholar 

  39. Madrid LV, Mayo MW, Reuther JY, Baldwin AS Jr (2001) Akt stimulates the transactivation potential of the RelA/p65 Subunit of NF-kappa B through utilization of the Ikappa B kinase and activation of the mitogen-activated protein kinase p38. J Biol Chem 276:18934–18940

    Article  PubMed  CAS  Google Scholar 

  40. Wellwood CRL, Cole RA (2004) Relevance of carnosic acid concentrations to the selection of Rosemary, Rosmarinus officinalis (L.), accessions for optimization of antioxidant yield. J Agric Food Chem 52:6101–6107

    Article  PubMed  CAS  Google Scholar 

  41. Yan H, Wang L, Li X et al (2009) High-performance liquid chromatography method for determination of carnosic acid in rat plasma and its application to pharmacokinetic study. Biomed Chromatogr 23:776–781

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Network Project (NWP 0038) and Supra Institutional Project (SIP 007) grant of the Council of Scientific and Industrial Research, Government of India.

Conflict of interest

The authors confirm that there are no conflicts of interest.

Author information

Authors and Affiliations

  1. Molecular Cell Biology Laboratory, Infectious Disease and Immunology Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Kolkata, 700032, India

    Susanta Kar, Shreyasi Palit, Writoban Basu Ball & Pijush K. Das

Authors
  1. Susanta Kar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shreyasi Palit
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Writoban Basu Ball
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Pijush K. Das
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pijush K. Das.

Additional information

Susanta Kar and Shreyasi Palit contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kar, S., Palit, S., Ball, W.B. et al. Carnosic acid modulates Akt/IKK/NF-κB signaling by PP2A and induces intrinsic and extrinsic pathway mediated apoptosis in human prostate carcinoma PC-3 cells. Apoptosis 17, 735–747 (2012). https://doi.org/10.1007/s10495-012-0715-4

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10495-012-0715-4

Keywords

Search

Navigation