International Journal of Clinical Oncology

, Volume 18, Issue 3, pp 380–388 | Cite as

Epigallocatechin-3-gallate potentiates curcumin’s ability to suppress uterine leiomyosarcoma cell growth and induce apoptosis

  • Akiko Kondo
  • Takashi TakedaEmail author
  • Bin Li
  • Kenji Tsuiji
  • Mari Kitamura
  • Tze Fang Wong
  • Nobuo Yaegashi
Original Article



Uterine leiomyosarcoma (LMS) has an unfavorable response to standard chemotherapeutic regimens. Two natural occurring compounds, curcumin and epigallocatechin gallate (EGCG), are reported to have anti-cancer activity. We previously reported that curcumin reduced uterine LMS cell proliferation by targeting the AKT–mTOR pathway. However, challenges remain in overcoming curcumin’s low bioavailability.


The human LMS cell line SKN was used. The effect of EGCG, curcumin or their combination on cell growth was detected by MTS assay. Their effect on AKT, mTOR, and S6 was detected by Western blotting. The induction of apoptosis was determined by Western blotting using cleaved-PARP specific antibody, caspase-3 activity and TUNEL assay. Intracellular curcumin level was determined by a spectrophotometric method. Antibody against EGCG cell surface receptor, 67-kDa laminin receptor (67LR), was used to investigate the role of the receptor in curcumin’s increased potency by EGCG.


In this study, we showed that the combination of EGCG and curcumin significantly reduced SKN cell proliferation more than either drug alone. The combination inhibited AKT, mTOR, and S6 phosphorylation, and induced apoptosis at a much lower curcumin concentration than previously reported. EGCG enhanced the incorporation of curcumin. 67LR antibody partially rescued cell proliferation suppression by the combination treatment, but was not involved in the EGCG-enhanced intracellular incorporation of curcumin.


EGCG significantly lowered the concentration of curcumin required to inhibit the AKT–mTOR pathway, reduce cell proliferation and induce apoptosis in uterine LMS cells by enhancing intracellular incorporation of curcumin, but the process was independent of 67LR.


Curcumin EGCG Uterine leiomyosarcoma mTOR AKT 67-kDa laminin receptor 



This work was supported, in part, by grants from the Japanese Ministry of Education, Science, Sports, and Culture, Tokyo, Japan (23592430).

Conflict of interest

The authors have no conflict of interest.


  1. 1.
    Leitao MM, Sonoda Y, Brennan MF et al (2003) Incidence of lymph node and ovarian metastases in leiomyosarcoma of the uterus. Gynecol Oncol 91:209–212PubMedCrossRefGoogle Scholar
  2. 2.
    Naaman Y, Shveiky D, Ben-Shachar I et al (2011) Uterine sarcoma: prognostic factors and treatment evaluation. Isr Med Assoc J 13:76–79PubMedGoogle Scholar
  3. 3.
    Muss HB, Bundy B, DiSaia PJ et al (1985) Treatment of recurrent or advanced uterine sarcoma: a randomized trial of doxorubicin versus doxorubicin and cyclophosphamide (a phase III trial of the Gynecologic Oncology Group). Cancer 55:1648–1653PubMedCrossRefGoogle Scholar
  4. 4.
    Omura GA, Major FJ, Blessing JA et al (1983) A randomized study of adriamycin with and without dimethyl triazenoimidazole carboxamide in advanced uterine sarcomas. Cancer 52:626–632PubMedCrossRefGoogle Scholar
  5. 5.
    Sutton G, Blessing JA, Malfetano JH (1996) Ifosfamide and doxorubicin in the treatment of advanced leiomyosarcomas of the uterus: a Gynecologic Oncology Group Study. Gynecol Oncol 62:226–229PubMedCrossRefGoogle Scholar
  6. 6.
    Piver MS, Lele SB, Marchetti DL et al (1988) Effect of adjuvant chemotherapy on time to recurrence and survival of stage I uterine sarcomas. J Surg Oncol 38:233–239PubMedCrossRefGoogle Scholar
  7. 7.
    Omura GA, Blessing JA, Major F et al (1985) A randomized clinical trial of adjuvant adriamycin in uterine sarcomas: a Gynecologic Oncology Group study. J Clin Oncol 3:1240–1245PubMedGoogle Scholar
  8. 8.
    Hernando E, Charytonowicz E, Dudas ME et al (2007) The AKT–mTOR pathway plays a critical role in the development of leiomyosarcomas. Nat Med 13:748–753PubMedCrossRefGoogle Scholar
  9. 9.
    Amant F, Coosemans A, Debiec-Rychter M et al (2009) Clinical management of uterine sarcomas. Lancet Oncol 10:1188–1198PubMedCrossRefGoogle Scholar
  10. 10.
    Sarbassov DD, Ali SM, Kim DH et al (2004) Rictor, a novel binding partner of mTOR, defines a rapamycin-insensitive and raptor-independent pathway that regulates the cytoskeleton. Curr Biol 14:1296–1302PubMedCrossRefGoogle Scholar
  11. 11.
    Breuleux M, Klopfenstein M, Stephan C et al (2009) Increased AKT S473 phosphorylation after mTORC1 inhibition is rictor dependent and does not predict tumor cell response to PI3K/mTOR inhibition. Mol Cancer Ther 8:742–753PubMedCrossRefGoogle Scholar
  12. 12.
    Sarbassov DD, Guertin DA, Ali SM et al (2005) Phosphorylation and regulation of Akt/PKB by the rictor–mTOR complex. Science 307:1098–1101PubMedCrossRefGoogle Scholar
  13. 13.
    Lambert JD, Yang CS (2003) Mechanisms of cancer prevention by tea constituents. J Nutr 133:3262S–3267SPubMedGoogle Scholar
  14. 14.
    Yang CS, Sang S, Lambert JD et al (2006) Possible mechanisms of the cancer-preventive activities of green tea. Mol Nutr Food Res 50:170–175PubMedCrossRefGoogle Scholar
  15. 15.
    Brown MD (1999) Green tea (Camellia sinensis) extract and its possible role in the prevention of cancer. Altern Med Rev 4:360–370PubMedGoogle Scholar
  16. 16.
    Bettuzzi S, Brausi M, Rizzi F et al (2006) Chemoprevention of human prostate cancer by oral administration of green tea catechins in volunteers with high-grade prostate intraepithelial neoplasia: a preliminary report from a one-year proof-of-principle study. Cancer Res 66:1234–1240PubMedCrossRefGoogle Scholar
  17. 17.
    Toda M, Okubo S, Ikigai H et al (1990) Antibacterial and anti-hemolysin activities of tea catechins and their structural relatives. Nippon Saikingaku Zassi 45:561–566CrossRefGoogle Scholar
  18. 18.
    Lin YL, Lin JK (1997) (−)-Epigallocatechin-3-gallate blocks the induction of nitric oxide synthase by down-regulating lipopolysaccharide-induced activity of transcription factor nuclear factor-κB. Mol Pharmacol 52:465–472PubMedGoogle Scholar
  19. 19.
    Maeda-Yamamoto M, Inagaki N, Kitaura J et al (2004) O-Methylated catechins from tea leaves inhibit multiple protein kinases in mast cells. J Immunol 172:4486–4492PubMedGoogle Scholar
  20. 20.
    Wheeler DS, Catravas JD, Odoms K et al (2004) Epigallocatechin-3-gallate, a green tea-derived polyphenol, inhibits IL-1β-dependent proinflammatory signal transduction in cultured respiratory epithelial cells. J Nutr 134:1039–1044PubMedGoogle Scholar
  21. 21.
    Yang F, Oz HS, Barve S et al (2001) The green tea polyphenol (−)-epigallocatechin-3-gallate blocks nuclear factor-κB activation by inhibiting IκB kinase activity in the intestinal epithelial cell line IEC-6. Mol Pharmacol 60:528–533PubMedGoogle Scholar
  22. 22.
    Van Aller GS, Carson JD, Tang W et al (2011) Epigallocatechin gallate (EGCG), a major component of green tea, is a dual phosphoinositide-3-kinase/mTOR inhibitor. Biochem Biophys Res Co 406:194–199CrossRefGoogle Scholar
  23. 23.
    Anand P, Sundaram C, Jhurani S et al (2008) Curcumin and cancer: an “old-age” disease with an “age-old” solution. Cancer Lett 267:133–164PubMedCrossRefGoogle Scholar
  24. 24.
    Wong TF, Takeda T, Li B et al (2011) Curcumin disrupts uterine leiomyosarcoma cells through AKT–mTOR pathway inhibition. Gynecol Oncol 122:141–148PubMedCrossRefGoogle Scholar
  25. 25.
    Somers-Edgar TJ, Scandlyn MJ, Stuart EC et al (2008) The combination of epigallocatechin gallate and curcumin suppresses ERα-breast cancer cell growth in vitro and in vivo. Int J Cancer 122:1966–1971PubMedCrossRefGoogle Scholar
  26. 26.
    Ghosh AK, Kay NE, Secreto CR et al (2009) Curcumin inhibits prosurvival pathways in chronic lymphocytic leukemia B cells and may overcome their stromal protection in combination with EGCG. Clin Cancer Res 15:1250–1258PubMedCrossRefGoogle Scholar
  27. 27.
    Saha A, Kuzuhara T, Echigo N et al (2010) New role of (−)-epicatechin in enhancing the induction of growth inhibition and apoptosis in human lung cancer cells by curcumin. Cancer Prev Res 3:953–962CrossRefGoogle Scholar
  28. 28.
    Tachibana H, Koga K, Fujimura Y et al (2004) A receptor of green tea polyphenol EGCG. Nat Struct Mol Biol 11:380–381PubMedCrossRefGoogle Scholar
  29. 29.
    Ishiwata I, Nozawa S, Nagal S et al (1977) Establishment of a human leiomyosarcoma cell line. Cancer Res 37:658–664PubMedGoogle Scholar
  30. 30.
    Byun EH, Fujimura Y, Yamada K et al (2010) TLR4 signaling inhibitory pathway induced by green tea polyphenol epigallocatechin-3-gallate through 67-kDa laminin receptor. J Immunol 185:33–45CrossRefGoogle Scholar
  31. 31.
    Holy EW, Stänpfli SF, Akhmedov A et al (2010) Laminin receptor activation inhibits endothelial tissue factor expression. J Mol Cell Cardiol 48:1138–1145PubMedCrossRefGoogle Scholar
  32. 32.
    Kuesap J, Li B, Satarug S et al (2008) Prostaglandin D2 induces heme oxygenase-1 in human retinal pigment epithelial cells. Biochem Biophys Res Commun 367:413–419PubMedCrossRefGoogle Scholar
  33. 33.
    Scott DW, Loo G (2007) Curcumin-induced GADD153 upregulation: modulation by glutathione. J Cell Biochem 101:307–320PubMedCrossRefGoogle Scholar
  34. 34.
    Shoba G, Joy D, Joseph T et al (1998) Influence of piperine on the pharmacokinetics of curcumin in animals and human volunteers. Planta Med 64:353–356PubMedCrossRefGoogle Scholar
  35. 35.
    Telang N, Katdare M (2007) Combinatorial prevention of carcinogenic risk in a model for familial colon cancer. Oncol Rep 17:909–914PubMedGoogle Scholar
  36. 36.
    Wan X, Harkavy B, Shen N et al (2007) Rapamycin induces feedback activation of Akt signaling through an IGF-1R-dependent mechanism. Oncogene 26:1932–1940PubMedCrossRefGoogle Scholar
  37. 37.
    Tewari M, Quan LT, O’Rourke K et al (1995) Yama/CPP32β, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose)polymerase. Cell 81:801–809PubMedCrossRefGoogle Scholar
  38. 38.
    Nagata S (2000) Apoptotic DNA fragmentation. Exp Cell Res 256:12–18PubMedCrossRefGoogle Scholar

Copyright information

© Japan Society of Clinical Oncology 2012

Authors and Affiliations

  • Akiko Kondo
    • 1
  • Takashi Takeda
    • 1
    • 2
    Email author
  • Bin Li
    • 2
  • Kenji Tsuiji
    • 2
  • Mari Kitamura
    • 1
  • Tze Fang Wong
    • 1
  • Nobuo Yaegashi
    • 1
    • 2
  1. 1.Department of Obstetrics and GynecologyTohoku University Graduate School of MedicineSendaiJapan
  2. 2.Department of Traditional Asian MedicineTohoku University Graduate School of MedicineSendaiJapan

Personalised recommendations