Skip to main content
NCBI home page
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 2000 Jan;91(1):34–40. doi: 10.1111/j.1349-7006.2000.tb00857.x

Green Tea Polyphenols Induce Apoptosis in vitro in Peripheral Blood T Lymphocytes of Adult T‐Cell Leukemia Patients

Hong‐Chuan Li 1, Shinji Yashiki 1, Junichiro Sonoda 2, Hong Lou 1, Subrata K Ghosh 3, John J Byrnes 3, Carolina Lema 1, Toshinobu Fujiyoshi 1, Mitsuaki Karasuyama 4, Shunro Sonoda 1,✉
PMCID: PMC5926221  PMID: 10744042

Abstract

Green tea polyphenols (TEA) are known to exhibit antioxidative activity as well as tumor‐suppressing activity. In order to examine the tumor‐suppressing activity of TEA against adult T‐cell leukemia (ATL), we cultivated peripheral blood T lymphocytes of ATL patients (ATL PBLs), an HTLV‐I‐infected T‐cell line (KODV) and healthy controls (normal PBLs) for 3 days in the presence of TEA and its main constituent, epigallocatechin‐3‐gallate (EGCg), to measure cell proliferation and apoptosis, and to quantitate mRNAs of HTLV‐I pX and β‐actin genes of the cultured cells. Growth of ATL PBLs was significantly inhibited by 9–27 μg/ml of TEA and EGCg, in contrast to minimal growth inhibition of T cells of normal PBLs. Inhibition of KODV was intermediate between ATL PBLs and normal PBLs. The ATL PBLs and KODV treated with 27 μg/ml of either TEA or EGCg induced apoptotic DNA fragmentation, producing terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labeling (TUNEL)‐positive cells, while the normal PBLs treated with the same concentration of TEA or EGCg produced a negligibly small number of TUNEL‐positive cells, in which apoptotic DNA fragmentation was not detectable. Expression of HTLV‐I pX mRNA was suppressed more than 90% in ATL PBLs by treatment with 3–27 μg/ml of either TEA or EGCg, while expression of β‐actin mRNA was much less suppressed by treatment with the same concentration of TEA or EGCg. These results indicate that TEA and EGCg inhibit growth of ATL PBLs, as well as HTLV‐I‐infected T‐cells, by suppressing HTLV‐I pX gene expression and inducing apoptotic cell death.

Keywords: EGCg, HTLV‐I, ATL, Apoptosis, Green tea

Full Text

The Full Text of this article is available as a PDF (293.1 KB).

REFERENCES

  • 1. ) Katiyar , S. K. , Mohan , R. R. , Agarwal , R. and Mukhtar , H.Protection against induction of mouse skin papillomas with low and high risk of conversion to malignancy by green tea polyphenols . Carcinogenesis , 18 , 497 – 502 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) Asano , Y. , Okamura , S. , Ogo , T. , Eto , T. , Otsuka , T. and Niho , Y.Effect of (‐)‐epigallocatechin gallate on leukemic blast cells from patients with acute myeloblastic leukemia . Life Sci. , 60 , 135 – 142 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 3. ) Valcic , S. , Timmermann , B. N. , Alberts , D. S. , Wächter , G. A. , Krutzsch , M. , Wymer , J. and Guillén , J. M.Inhibitory effect of six green tea catechins and caffeine on the growth of four selected human tumor cell lines . Anticancer Drugs , 7 , 461 – 468 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 4. ) Hibasami , H. , Komiya , T. , Achiwa , Y. , Ohnishi , K. , Kojima , T. , Nakanishi , K. , Akashi , K. and Hara , Y.Induction of apoptosis in human stomach cancer cells by green tea catechins . Oncol. Rep. , 5 , 527 – 529 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Hibasami , H. , Achiwa , Y. , Fujikawa , T. and Komiya , T.Induction of programmed cell death (apoptosis) in human lymphoid leukemia cells by catechin compounds . Anticancer Res. , 16 , 1943 – 1946 ( 1996. ). [PubMed] [Google Scholar]
  • 6. ) Zhao , Y. , Cao , J. , Ma , H. and Liu , J.Apoptosis induced by tea polyphenols in HL‐60 cells . Cancer Lett. , 121 , 163 – 167 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Ahmad , N. , Feyes , D. K. , Nieminen , A. L. , Agarwal , R. and Mukhtar , H.Green tea constituent epigallocatechin‐3‐gallate and induction of apoptosis and cell cycle arrest in human carcinoma cells . J. Natl. Cancer Inst. , 89 , 1881 – 1886 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Paschka , A. G. , Butler , R. and Young , C. Y. F.Induction of apoptosis in prostate cancer cell lines by the green tea component, (‐)‐epigallocatechin‐3‐gallate . Cancer Lett. , 130 , 1 – 7 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Wyllie , A. H. , Kerr , J. F. R. and Currie , A. R.Cell death. The significance of apoptosis . Int. Rev. Cytol. , 68 , 251 – 306 ( 1980. ). [DOI] [PubMed] [Google Scholar]
  • 10. ) Suganuma , M. , Okabe , S. , Sueoka , E. , Iida , N. , Komori , A. , Kim , S. J. and Fujiki , H.A new process of cancer prevention mediated through inhibition of tumor necrosis factor α expression . Cancer Res. , 15 , 3711 – 3715 ( 1996. ). [PubMed] [Google Scholar]
  • 11. ) Lu , L. H. , Lee , S. S. and Huang , H. C.Epigallocatechin suppression of proliferation of vascular smooth muscle cells: correlation with c‐jun and JNK . Br. J. Pharmacol. , 124 , 1227 – 1237 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. ) Chan , M. M. Y. , Fong , D. , Ho , C. T. and Huang , H. I.Inhibition of inducible nitric oxide synthase gene expression and enzyme activity by epigallocatechin gallate, a natural product from green tea . Biochem. Pharmacol. , 54 , 1281 – 1286 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Katiyar , S. K. , Rupp , C. O. , Korman , N. J. , Agarwal , R. and Mukhtar , H.Inhibition of 12‐O‐tetradecanoylphorbol‐13‐acetate and other skin tumor‐promoter‐caused induction of epidermal interleukin‐1 α mRNA and protein expression in SENCAR mice by green tea polyphenols . J. Invest. Dermatol. , 105 , 394 – 398 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 14. ) Wang , Z. Y. , Cheng , S. J. , Zhou , Z. C. , Athar , M. , Khan , W. A. , Bickers , D. R. and Mukhtar , H.Antimutagenic activity of green tea polyphenols . Mutat. Res. , 223 , 273 – 285 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Katiyar , S. K. and Mukhtar , H.Tea antioxidants in cancer chemoprevention . J. Cell. Biochem. , 27 ( Suppl. ), S59 – S67 ( 1997. ). [PubMed] [Google Scholar]
  • 16. ) Nakachi , K. , Suemasu , K. , Suga , K. , Takeo , T. , Imai , K. and Higashi , Y.Influence of drinking green tea on breast cancer malignancy among Japanese patients . Jpn. J. Cancer Res. , 89 , 254 – 261 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17. ) Inoue , M. , Tajima , K. , Hirose , K. , Hamajima , N. , Takezaki , T. , Kuroishi , T. and Tominaga , S.Tea and coffee consumption and the risk of digestive tract cancers: data from a comparative case‐referent study in Japan . Cancer Causes Control , 9 , 209 – 216 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Okabe , S. , Ochiai , Y. , Aida , M. , Park , K. , Kim , S.‐J. , Nomura , T. , Suganuma , M. and Fujiki , H.Mechanistic aspects of green tea as a cancer preventive: effect of components on human stomach cancer cell lines . Jpn. J. Cancer Res. , 90 , 733 – 739 ( 1999. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. ) Takatsuki , K. , Uchiyama , T. , Sagawa , K. and Yodoi , J.Adult T cell leukemia in Japan . In “ Topics in Hematology ,” ed. Seno S. , Takaku F. and Irino S. , pp. 73 – 77 ( 1977. ). Excerpta Medica; , Amsterdam . [Google Scholar]
  • 20. ) Uchiyama , T. , Yodoi , J. , Sagawa , K. , Takatsuki , K. and Uchino , H.Adult T‐cell leukemia: clinical and hematologic features of 16 cases . Blood , 50 , 481 – 492 ( 1977. ). [PubMed] [Google Scholar]
  • 21. ) Poiesz , B. J. , Ruscetti , F. W. , Gazdar , A. F. , Bunn , P. A. , Minna , J. D. and Gallo , R. C.Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T‐cell lymphoma . Proc. Natl. Acad. Sci. USA , 77 , 7415 – 7419 ( 1980. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22. ) Hinuma , Y. , Nagata , K. , Hanaoka , M. , Nakai , M. , Matsumoto , T. , Kinoshita , K. I , Shirakawa , S. and Miyoshi , I.Adult T‐cell leukemia: antigen in an ATL cell line and detection of antibodies to the antigen in human sera . Proc. Natl. Acad. Sci. USA , 78 , 6476 – 6480 ( 1981. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. ) Yoshida , M. , Miyoshi , I. and Hinuma , Y.Isolation and characterization of retrovirus from cell lines of human adult T‐cell leukemia and its implication in the disease . Proc. Natl. Acad. Sci. USA , 79 , 2031 – 2035 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. ) Tajima , K. , Tominaga , S. , Suchi , T. , Fukui , H. , Komoda , H. and Hinuma , Y.HTLV‐I carriers among migrants from an ATL‐endemic area to ATL non‐endemic metropolitan areas in Japan . Int. J. Cancer , 37 , 383 – 387 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 25. ) Blattner , W. A. , Kalyanaraman , V. S. , Robert‐Guroff , M. , Lister , T. A. , Galton , D. A. G , Sarin , P. S. , Crawford , M. H. , Catovsky , D. , Greaves , M. and Gallo , R. C.The human type‐C retrovirus, HTLV, in Blacks from the Caribbean region, and relationship to adult T‐cell leukemia/lymphoma . Int. J. Cancer , 30 , 257 – 264 ( 1982. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) Seiki , M. , Hikikoshi , A. , Taniguchi , T. and Yoshida , M.Expression of the pX gene of HTLV‐I: general splicing mechanism in the HTLV family . Science , 228 , 1532 – 1534 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 27. ) Ninomiya , M. , Unten , L. and Kim , M.Chemical and physicochemical properties of green tea polyphenols . In “ Chemistry and Applications of Green Tea ,” ed. Yamamoto T. , Juneja L. R. , Chu D. C. and Kim M. , pp. 23 – 35 ( 1997. ). CRC Press LLC; , Boca Raton, New York . [Google Scholar]
  • 28. ) Nakagawa , K. and Miyazawa , T.Chemiluminescence‐highperformance liquid chromatographic determination of tea catechin, (‐)‐epigallocatechin 3‐gallate, at picomole levels in rat and human plasma . Anal. Biochem. , 248 , 41 – 49 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 29. ) Osame , M. , Igata , A. and Matsumoto , M.HTLV‐I‐associated myelopathy (HAM) revisited . In “ HTLV‐I and the Nervous System ,” ed. Roman G. C. , Vernant J. C. and Osame M. , pp. 213 – 223 ( 1989. ). Alan R. Liss, Inc. , New York . [Google Scholar]
  • 30. ) Katahira , Y. , Yashiki , S. , Fujiyoshi , T. , Nomura , K. , Tara , M. , Mori , M. , Setoyama , M. , Kanzaki , T. , Shida , H. and Sonoda , S.In vitro, induction of cytotoxic T lymphocytes against HTLV‐I‐infected T‐cells from adult T‐cell leukemia patients, asymptomatic HTLV‐I carriers and seronegative healthy donors . Jpn. J. Cancer Res. , 86 , 21 – 27 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31. ) Ishiyama , M. , Miyazono , Y. , Sasamoto , K. , Ohkura , Y. and Ueno , K.A highly water‐soluble disulfonated tetrazolium salt as a chromogenic indicator for NADH as well as cell viability . Talanta , 44 , 1299 – 1305 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 32. ) Ioannou , Y. A. and Chen , F. W.Quantitation of DNA fragmentation in apoptosis . Nucleic Acids Res. , 24 , 992 – 993 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33. ) Herrmann , M. , Lorenz , H. M. , Voll , R. , Grünke , M. , Woith , W. and Kalden , J. R.A rapid and simple method for the isolation of apoptotic DNA fragments . Nucleic Acids Res. , 22 , 5506 – 5507 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34. ) Gavrieli , Y. , Sherman , Y. and Ben‐Sasson , S. A.Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation . J. Cell Biol. , 119 , 493 – 501 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35. ) Higashiyama , Y. , Katamine , S. , Kohno , S. , Mukae , H. , Hino , S. , Miyamoto , T. and Hara , K.Expression of human T lymphotropic virus type 1 (HTLV‐1) tax/rex gene in fresh bronchoalveolar lavage cells of HTLV‐1‐infected individuals . Clin. Exp. Immunol. , 96 , 193 – 201 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36. ) Kinoshita , T. , Shimoyama , M. , Tobinai , K. , Ito , M. , Ito , S. I. , Ikeda , S. , Tajima , K. , Shimotohno , K. and Sugimura , T.Detection of mRNA for the tax1/rex1 gene of human T‐cell leukemia virus type I in fresh peripheral blood mononuclear cells of adult T‐cell leukemia patients and viral carriers by using the polymerase chain reaction . Proc. Natl. Acad. Sci. USA , 86 , 5620 – 5624 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37. ) Ponte , P. , Ng , S. Y. , Engel , J. , Gunning , P. and Kedes , L.Evolutionary conservation in the untranslated regions of actin mRNAs: DNA sequence of a human beta‐actin cDNA . Nucleic Acids Res. , 12 , 1687 – 1696 ( 1984. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 38. ) Chu , D. C. and Juneja , L. R.General chemical composition of green tea and its infusion . In “ Chemistry and Applications of Green Tea ,” ed. Yamamoto T. , Juneja L. R. , Chu D. C. and Kim M. , pp. 13 – 22 ( 1997. ). CRC Press LLC; , Boca Raton, New York . [Google Scholar]
  • 39. ) Heta , T. , Fujimoto , T. , Tsushima , H. , Murata , K. , Tsukasaki , K. , Atogami , S. , Sohda , H. , Honda , S. , Mine , M. , Yamada , Y. , Ikeda , S. , Kamihira , S. and Tomonaga , M.Multi‐clonal expansion of unique human T‐lymphotropic virus type‐I‐infected T cells with high growth potential in response to interleukin‐2 in prodromal phase of adult T cell leukemia . Leukemia , 13 , 215 – 221 ( 1999. ). [DOI] [PubMed] [Google Scholar]
  • 40. ) Uchiyama , T. , Hori , T. , Tsudo , M. , Wano , Y. , Umadome , H. , Tamori , S. , Yodoi , J. , Maeda , M. , Sawami , H. and Uchino , H.Interleukin‐2 receptor (Tac antigen) expressed on adult T cell leukemia cells . J. Clin. Invest. , 76 , 446 – 453 ( 1985. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41. ) Lee , M. J. , Wang , Z. Y. , Li , H. , Chen , L. , Sun , Y. , Gobbo , S. , Balentine , D. A. and Yang , C. S.Analysis of plasma and urinary tea polyphenols in human subjects . Cancer Epidemiol. Biomarkers Prev. , 4 , 393 – 399 ( 1995. ). [PubMed] [Google Scholar]
  • 42. ) Nakagawa , K. , Okuda , S. and Miyazawa , T.Dose‐dependent incorporation of tea catechins, (‐)‐epigallocatechin‐3‐gallate and (‐)‐epigallocatechin, into human plasma . Biosci. Biotechnol. Biochem. , 61 , 1981 – 1985 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 43. ) Suga , K. , Imai , K. , Sueoka , N. and Nakachi , K.Phase I clinical trial with green tea tablets in a Japanese healthy population . Cancer Pre. Intern. , 3 , 79 – 88 ( 1998. ). [Google Scholar]
  • 44. ) Fujiki , H. , Suganuma , M. , Okabe , S. , Sueoka , E. , Suga , K. , Imai , K. , Nakachi , K. and Kimura , S.Mechanistic findings of green tea as cancer preventive for humans . Proc. Soc. Exp. Biol. Med. , 220 , 225 – 228 ( 1999. ). [DOI] [PubMed] [Google Scholar]
  • 45. ) Austin , C. A. , Patel , S. , Ono , K. , Nakane , H. and Fisher , M.Site‐specific DNA cleavage by mammalian DNA topoisomerase II induced by novel flavone and catechin derivatives . Biochem. J. , 282 , 883 – 889 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 46. ) Jankun , J. , Selman , S. H. , Swiercz , R. and Skrzypczak‐Jankun , E.Why drinking green tea could prevent cancer . Nature , 387 , 561 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 47. ) Dong , Z. , Ma , W. , Huang , C. and Yang , C. S.Inhibition of tumor promoter‐induced activator protein 1 activation and cell transformation by tea polyphenols, (‐)‐epigallocatechin gallate and theaflavins . Cancer Res. , 57 , 4414 – 4419 ( 1997. ). [PubMed] [Google Scholar]
  • 48. ) Cao , Y. and Cao , R.Angiogenesis inhibited by drinking tea . Nature , 398381 , ( 1999. ). [DOI] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES