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Caffeic acid phenethyl ester triggers apoptosis through induction of loss of mitochondrial membrane potential in CCRF-CEM cells

  • Çığır Biray Avcı
  • Cumhur Gündüz
  • Yusuf Baran
  • Fahri Şahin
  • Sunde Yılmaz
  • Zeynep Ozlem Dogan
  • Güray Saydam
Original Paper

Abstract

Purpose

CAPE (caffeic acid phenethyl ester) is one of the most valuable and investigated component of propolis which is composed by honeybees. In the current study, we aimed at examining apoptotic effects of CAPE on CCRF-CEM leukemic cells and at determining the roles of mitochondrial membrane potential (MMP) in cell death.

Methods

Trypan blue and XTT methods were used to evaluate the cytotoxicity. Apoptosis was examined by ELISA-based oligonucleotide and acridine orange/ethidium bromide dye techniques. Loss of mitochondrial membrane potential was evaluated using JC-1 dye by flow cytometric analysis and under fluorescent microscope.

Results

We detected the time- and dose-dependent increases in cytotoxic effect of CAPE on CCRF-CEM cells. ELISA and acridine orange/ethidium bromide results showed that apoptotic cell population increased significantly in CCRF-CEM cells exposed to increasing concentrations of CAPE. On the other hand, there was significant loss of MMP determined in response to CAPE in CCRF-CEM cells.

Conclusion

This in vitro data by being supported with clinical data may open the way of the potential use of CAPE for the treatment of leukemia.

Keywords

CAPE ALL Apoptosis Mitochondrial membrane potential Cytotoxicity 

References

  1. Asoh S, Mori T, Hayashi J, Ohta S (1996) Expression of the apoptosis-mediator Fas is enhanced by dysfunctional mitochondria. J Biochem 120:600–607PubMedGoogle Scholar
  2. Baran Y, Salas A, Senkal CE, Bielawski J, Gunduz U, Obeid LM, Ogretmen B (2007a) Alterations of human longevity assurance gene 1 (LASS1)/sphingosine kinase-1-dependent ceramide generation and metabolism involve in the regulation of imatinib-induced apoptosis and resistance in K562 human chronic myeloid leukemia (CML) cells. J Biol Chem 282(15):10922–10934CrossRefPubMedGoogle Scholar
  3. Baran Y, Gur B, Ural AU, Avcu F, Gunduz U (2007b) Upregulation of multi drug resistance genes in doxorubicin resistant human acute myelogenous leukemia cells and reversal of the resistance. Hematology 12(6):511–517CrossRefPubMedGoogle Scholar
  4. Baran Y, Ural AU, Gunduz U (2007c) Mechanisms of cellular resistance to imatinib in human chronic myeloid leukemia cells. Hematology 12(6):497–503CrossRefPubMedGoogle Scholar
  5. Baxa DM, Luo X, Yoshimura FK (2005) Genistein Induces apoptosis in T lymphoma cells via mitochondrial damage. Nutr Cancer 51:93–101CrossRefPubMedGoogle Scholar
  6. Berger N, Ben Bassat H, Klein BY, Laskov R (1992) Cytotoxicity of NF-kappaB inhibitors Bay 11–7085 and caffeic acid phenethyl ester to Ramos and other human B-lymphoma cell lines. Exp Hematol 35:1495–1450CrossRefGoogle Scholar
  7. Bhimani RS, Troll W, Grunberger D, Frenkel K (1993) Inhibition of oxidative stress in HeLa cells by chemopreventive agents. Cancer Res 53:4528–4533PubMedGoogle Scholar
  8. Borrelli F, Izzo AA, Di Carlo G, Maffia P, Russo A, Maiello FM, Capasso F, Mascolo N (2002) Effect of a propolis extract and cafeic acid phenethyl ester on formation of aberrant crypt foci and tumors in the rat colon. Fitoterapia 73:38–43CrossRefGoogle Scholar
  9. Burke TR, Fesen MR, Mazumder A, Wang J, Carothers AM, Grunberger D, Driscoll J, Kohn K, Pommier Y (1995) Hydroxylated aromatic inhibitors of HIV-1 integrase. J Med Chem 38:4171–4178CrossRefPubMedGoogle Scholar
  10. Cassileth B, Yeung KS, Gubili J (2008) Herbs and other botanicals in cancer patient care. Curr Treat Options Oncol 9(2–3):109–116CrossRefPubMedGoogle Scholar
  11. Chen JH, Shao Y, Huang MT, Chin CK, Ho CT (1996) Inhibitory effect of caffeic acid phenethyl ester on human leukemia HL-60 cells. Cancer Lett 108(2):211–214CrossRefPubMedGoogle Scholar
  12. Chen Y, Shiao M, Wang SY (2001) The antioxidant caffeic acid phenethyl ester induces apoptosis associated with selective scavenging of hydrogen peroxide in human leukemic HL-60 cells. Anti Cancer Drugs 12:143–149CrossRefPubMedGoogle Scholar
  13. Chen JC, Zhang X, Singleton TP, Kiechle FL (2004) Mitochondrial membrane potential change induced by Hoechst 33342 in myelogenous leukemia cell line HL-60. Ann Clin Lab Sci 34:458–466PubMedGoogle Scholar
  14. Chen YJ, Liao HF, Tsai TH, Wang SY, Shiao MS (2005) Caffeic acid phenethyl ester preferentially sensitizes CT26 colorectal adenocarcinoma to ionizing radiation without affecting bone marrow radioresponse. Int J Radiat Oncol Biol Phys 63:1252–1261CrossRefPubMedGoogle Scholar
  15. Chen MJ, Chang WH, Lin CC, Liu CY, Wang TE, Chu CH, Shih SC, Chen YJ (2008) Caffeic acid phenethyl ester induces apoptosis of human pancreatic cancer cells involving caspase and mitochondrial dysfunction. Pancreatology 8:566–576CrossRefPubMedGoogle Scholar
  16. Chiao C, Carothers AM, Grunberger D, Solomon G, Preston GA, Barrett JC (1995) Apoptosis and altered redox state induced by caffeic acid phenethyl ester (CAPE) in transformed rat fibroblast cells. Cancer Res 55:3576–3583PubMedGoogle Scholar
  17. Chung TW, Moon SK, Chang YC, Ko JH, Lee YC, Cho G, Kim SH, Kim JG, Kim CH (2004) Novel and therapeutic effect of caffeic acid and caffeic acid phenyl ester on hepatocarcinoma cells: complete regression of hepatoma growth and metastasis by dual mechanism. FASEB J 18:1670–1681CrossRefPubMedGoogle Scholar
  18. Cossarizza A, Baccarani-Contri M, Kalashnikova G, Franceschi C (1993) A new method for the cytofluorimetric analysis of mitochondrial membrane potential using the J-aggregate forming lipophilic cation 5, 50, 6, 60-tetrachloro-1, 10, 3, 30 tetraethylbenzimidazolcarbocyanine iodide (JC-1). Biochem Biophys Res Commun 197:40–45CrossRefPubMedGoogle Scholar
  19. Crazzolara R, Bendall L (1999) Emerging treatments in acute lymphoblastic leukemia. Curr Cancer Drug Targets 9:19–31CrossRefGoogle Scholar
  20. Demestre M, Messerli SM, Celli N, Shahhossini M, Kluwe L, Mautner V, Maruta H (2009) CAPE (caffeic acid phenethyl ester)-based propolis extract (Bio 30) suppresses the growth of human neurofibromatosis (NF) tumor xenografts in mice. Phytother Res 23:226–230CrossRefPubMedGoogle Scholar
  21. Dörrie J, Gerauer H, Wachter Y, Zunino SJ (2001) Resveratrol induces extensive apoptosis by depolarizing mitochondrial membranes and activating caspase-9 in acute lymphoblastic leukemia cells. Cancer Res 61:4731–4739PubMedGoogle Scholar
  22. Engdal S, Klepp O, Nilsen OG (2009) Identification and exploration of herb-drug combinations used by cancer patients. Integr Cancer Ther 8(1):29–36CrossRefPubMedGoogle Scholar
  23. Fesen MR, Pommier Y, Leteurtre F, Hiroguchi S, Yung J, Kohn KW (1994) Inhibition of HIV-1 integrase by flavones, caffeic acid phenethyl ester (CAPE) and related compounds. Biochem Pharmacol 48:595–608CrossRefPubMedGoogle Scholar
  24. Gogvadze V, Orrenius S, Zhivotovsky B (2006) Multiple pathways of cytochrome c release from mitochondria in apoptosis. Biochim Biophys Acta 1757:639–647CrossRefPubMedGoogle Scholar
  25. Gogvadze V, Orrenius S, Zhivotovsky B (2008) Mitochondria in cancer cells: what is so special about them? Trends Cell Biol 18:165–173CrossRefPubMedGoogle Scholar
  26. Grunberger D, Banerjee R, Eisinger K, Oltz EM, Efros L, Caldwell M, Estevez V, Nakanishi K (1998) Preferential cytotoxicity on tumor cells by caffeic acid phenethyl ester isolated from propolis. Experientia 44:230–232CrossRefGoogle Scholar
  27. He YJ, Liu BH, Xiang DB, Qiao ZY, Fu T, He YH (2006) Inhibitory effect of caffeic acid phenethyl ester on the growth of SW480 colorectal tumor cells involves beta-catenin associated signaling pathway downregulation. World J Gastroenterol 12:4981–4985PubMedGoogle Scholar
  28. Hung MW, Shiao MS, Tsai LC, Chang GG, Chang TC (2003) Apoptotic effect of caffeic acid phenethyl ester and its ester and amide analogues in human cervical cancer ME180 cells. Anticancer Res 23:4773–4780PubMedGoogle Scholar
  29. Lee YJ, Liao PH, Chen WK, Yang CY (2000) Preferential cytotoxicity of caffeic acid phenethyl ester analogues on oral cancer cells. Cancer Lett 153:51–56CrossRefPubMedGoogle Scholar
  30. Lee YJ, Kuo HC, Chu CY, Wang CJ, Lin WC, Tseng TH (2003) Involvement of tumor suppressor protein p53 and p38 MAPK in caffeic acid phenethyl ester-induced apoptosis of C6 glioma cells. Biochem Pharmacol 66:2281–2289CrossRefPubMedGoogle Scholar
  31. Li D, Saldeen T, Romeo F, Mehta JL (2000) Oxidized LDL upregulates angiotensin II type 1 receptor expression in cultured human coronary artery endothelial cells: the potential role of transcription factor NF-kappaB. Circulation 102:1970–1976PubMedGoogle Scholar
  32. Liao H, Chen Y, Liu J (1999) Inhibitory effect of caffeic acid phenyl ester on angiogenesis, tumor invasion, and metastasis. J Agric Food Chem 51:7907–7912CrossRefGoogle Scholar
  33. Michaluart P, Masferrer JL, Carothers AM, Subbaramaiah K, Zweifel BS, Koboldt C, Mestre JR, Grunberger D, Sacks PG, Tanabe T, Dannenberg AJ (1999) Inhibitory effects of caffeic acid phenethyl ester on the activity and expression of cyclooxygenase-2 in human oral epithelial cells and in a rat model of inflammation. Cancer Res 59:2347–2352PubMedGoogle Scholar
  34. Neuzil J, Weber T, Gellert N, Weber C (2001) Selective cancer cell killing by alpha-tocopheryl succinate. Br J Cancer 84(1):87–89CrossRefPubMedGoogle Scholar
  35. Neuzil J, Wang XF, Dong LF, Low P, Ralph SJ (2006) Molecular mechanism of ‘mitocan’-induced apoptosis in cancer cells epitomizes the multiple roles of reactive oxygen species and Bcl-2 family proteins. FEBS Lett 5(22):5125–5129CrossRefGoogle Scholar
  36. Neuzil J, Dyason JC, Freeman R, Dong LF, Prochazka L, Wang XF, Scheffler I, Ralph SJ (2007) Mitocans as anti-cancer agents targeting mitochondria: lessons from studies with vitamin E analogues, inhibitors of complex II. J Bioenerg Biomembr 39:7265–7282CrossRefGoogle Scholar
  37. Orban Z, Mitsiades N, Burke TR Jr, Tsokos M, Chrousos GP (2000) Caffeic acid phenethyl ester induces leukocyte apoptosis, modulates nuclear factor-kappa B and suppresses acute inflammation. Neuroimmunomodulation 7:99–105CrossRefPubMedGoogle Scholar
  38. Rao CV, Desai D, Kaul B, Amin S, Reddy BS (1992) Effect of caffeic acid esters on carcinogen-induced mutagenicity and human colon adenocarcinoma cell growth. Chem Biol Interact 84(3):277–290CrossRefPubMedGoogle Scholar
  39. Sahin F, Avci CB, Avcu F, Ural AU, Sarper M, Omay SB, Saydam G (2007) Red grape seed extract and its compound resveratrol exert cytotoxic effect to various human cancer lines. Turk J Hematolol 24:102–109Google Scholar
  40. Salvioli S, Ardizzoni A, Franceschi C, Cossarizza A (1997) JC-1, but not DiOC6(3) or rhodamine 123, is a reliable fluorescent probe to assess Dc changes in intact cells:Implications for studies on mitochondrial functionality during apoptosis. FEBS Lett 411:77–82CrossRefPubMedGoogle Scholar
  41. Schmidt-Mende J, Gogvadze V, Hellström-Lindberg E, Zhivotovsky B (2006) Early mitochondrial alterations in ATRA-induced cell death. Cell Death Differ 13:119–128CrossRefPubMedGoogle Scholar
  42. Son S, Lewis BA (2002) Free radical scavenging and antioxidative activity of cafeic acid amide and ester analogues: structure activity relationship. J Agric Food Chem 50:468–472CrossRefPubMedGoogle Scholar
  43. Sud’ina GF, Mirzoeva OK, Pushkareva MA, Korshunova GA, Sumbatyan NV, Varfolomeev SD (1993) Caffeic acid phenethyl ester as a lipoxygenase inhibitor with antioxidant properties. FEBS Lett 329:21–24CrossRefPubMedGoogle Scholar
  44. Swerts K, de Moerloose B, Dhooge C, Noens L, Laureys G, Benoit Y, Philippé J (2004) Comparison of two functional flow cytometric assays to assess P-gp activity in acute leukemia. Leuk Lymphoma 45(11):2221–2228CrossRefPubMedGoogle Scholar
  45. Wang D, Xiang DB, He YJ, Li ZP, Wu XH, Mou JH, Xiao HL, Zhang QH (2005) Effect of caffeic acid phenethyl ester on proliferation and apoptosis of colorectal cancer cells in vitro. World J Gastroenterol 11:4008–4012PubMedGoogle Scholar
  46. Watabe M, Hishikawa K, Takayanagi A, Shimizu N, Nakaki T (2004) Caffeic acid phenethyl ester induces apoptosis by inhibition of NFkappaB and activation of Fas in human breast cancer MCF-7 cells. J Biol Chem 279:6017–6026CrossRefPubMedGoogle Scholar
  47. Wolvetang EJ, Johnson KL, Krauer K, Ralph SJ, Linnane AW (1994) Mitochondrial respiratory chain inhibitors induce apoptosis. FEBS Lett 339:40–44CrossRefPubMedGoogle Scholar
  48. Yu W, Sanders BG, Kline K (2003) RRR-alpha-tocopheryl succinate-induced apoptosis of human breast cancer cells involves Bax translocation to mitochondria. Cancer Res 63(10):2483–2491PubMedGoogle Scholar
  49. Zunino SJ, Storms DH (2006) Resveratrol-induced apoptosis is enhanced in acute lymphoblastic leukemia cells by modulation of the mitochondrial permeability transition pore. Cancer Lett 240:123–134CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Çığır Biray Avcı
    • 1
  • Cumhur Gündüz
    • 1
  • Yusuf Baran
    • 2
  • Fahri Şahin
    • 3
  • Sunde Yılmaz
    • 1
  • Zeynep Ozlem Dogan
    • 1
  • Güray Saydam
    • 3
  1. 1.Department of Medical Biology, School of MedicineEge UniversityBornovaTurkey
  2. 2.Department of Molecular Biology and GeneticsIzmir Institute of TechnologyUrlaTurkey
  3. 3.Department of Hematology, School of MedicineEge UniversityBornovaTurkey

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