Medical Oncology

, Volume 22, Issue 2, pp 129–138 | Cite as

In vitro and in vivo antitumorigenic activity of a mixture of lysine, proline, ascorbic acid, and green tea extract on human breast cancer lines MDA-MB-231 and MCF-7

  • M. Waheed Roomi
  • Vadim Ivanov
  • Tatiana Kalinovsky
  • Aleksandra Niedzwiecki
  • Matthias Rath
Original Article


Current treatments are generally ineffective once breast cancer has metastasized; median survival is reduced to 2–3 yr. Previous research studies demonstrating potent synergistic antitumor activity of lysine, proline, ascorbic acid, and epigallocatechin gallate prompted us to investigate the in vivo inhibitory effect of a nutrient mixture containing lysine, proline, arginine, ascorbic acid, and epigallocatechin gallate (NM) on the growth of human cancer xenografts in female athymic nude mice. Five to six week old female mice were inoculated with 3×106 breast cancer cells MDA-MB-231. After injection, the mice were randomly divided into two groups A and B; group A was fed a regular diet and group B with the regular diet supplemented with 0.5% of the nutrient mixture (NM). Four weeks later, the mice were sacrificed, and their tumors were excised, weighed, and processed for histology. We also tested the effect of NM in vitro on estrogen-receptor positive (ER+) MCF-7 and estrogen-receptor negative (ER) MDA-MB-231 breast cancer cell lines by measuring: cell proliferation by MTT assay, expression of MMPs by gelatinase zymography, invasion through Matrigel, and VEGF by ELISA. MCF-7 cells were also treated with estradiol to study enhanced invasion and expression of MMPs and VEGF. Results showed that NM inhibited the growth and reduced the size of tumors in female nude mice by 27%. Furthermore, histological evaluation revealed increased mitotic index, MMP-9 and VEGF secretion, and PAS material (mucin) in the control group tissues. In vitro studies showed NM inhibited MDA-MB-231 cell growth by 34% at 500 µg/mL and MCF-7 cell growth by 18% at 1000 µg/mL. Invasion of MDA-MB-231 through Matrigel was inhibited by 50%, 60%, and 95% by 10, 50, and 100 µg/mL of NM, respectively. The results of this study demonstrated that the nutrient mixture tested significantly suppressed tumor growth of breast cancer cells in female athymic nude mice and significantly inhibited MMP expression, angiogenesis, and invasion in breast cancer cells, in vitro, offering promise for therapeutic use in the treatment of breast cancer.

Key Words

Breast cancer MCF-7 MDA-MB-231 MMP VEGF Athymic nude mice xenograft Matrigel invasion antitumor 


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  1. 1.
    Ali SM, Harvey HA, Lipton A. Metastatic breast cancer: overview of treatment. Clin Orthop 2003; 415(Suppl): 132–137.Google Scholar
  2. 2.
    Pantel K, Muller V, Auer M, Nusser N, Harbeck N, Braun S. Detection and clinical implications of early systemic tumor cell dissemination in breast cancer. Clin Cancer Res 2003; 17:6326–6334.Google Scholar
  3. 3.
    Rath M, Pauling L. Plasmin-induced proteolysis and the role of apoprotein(a), lysine and synthetic analogs. Orthomol Med 1992; 7:17–23.Google Scholar
  4. 4.
    Netke SP, Roomi MW, Ivanov V, Niedzwiecki A, Rath M. A specific combination of ascorbic acid, lysine, proline and epigallocatechin gallate inhibits proliferation and extracellular matrix invasion of various human cancer cell lines. Res Commun Pharmacol Toxicol Emerging Drugs 2003; 2:37–50.Google Scholar
  5. 5.
    Skobe M, et al. Induction of tumor lymphangiogenesis by VEGF-C promotes breast cancer metastasis. Nat Med 2001; 7(2):192–198.PubMedCrossRefGoogle Scholar
  6. 6.
    Garvin S., Dabrosin C. Tamoxifen inhibits secretion of vascular endothelial growth factor in breast cancer in vivo. Cancer Res 2003; 24:8742–8748.Google Scholar
  7. 7.
    Bachmeier BE, Nerlich AG, Lichtinghagen R, Sommerhoff CP. Matrix metalloproteinases (MMPs) in breast cancer cell lines of different tumorigenicity. Anticancer Res 2001; 6A:3821–3828.Google Scholar
  8. 8.
    Koh WS, et al. Differential effects and transport kinetics of ascorbate derivatives in leukemic cell lines. Anticancer Res 1998; 8:2487–2493.Google Scholar
  9. 9.
    Roomi MW, House D, Eckert-Maksic M, Maksic ZB, Tsao CS. Growth suppression of malignant leukemia cell line in vitro by ascorbic acid (vitamin C) and its derivatives. Cancer Lett 1998; 122:93–99.PubMedCrossRefGoogle Scholar
  10. 10.
    Naidu KA, Karl RC, Naidu KA, Coppola D. Antiproliferative and proapoptotic effect of ascorbyl stearate in human pancreatic cancer cells: association with decreased expression of insulin-like growth factor 1 receptor. Dig Dis Sci 2003; 48(1):230–237.PubMedCrossRefGoogle Scholar
  11. 11.
    Anthony HM, Schorah CJ. Severe hypovitaminosis C in lung-cancer patients: the utilization of vitamin C in surgical repair and lymphocyte related host resistance. Br J Cancer 1982; 46:354–367.PubMedCrossRefGoogle Scholar
  12. 12.
    Nunez C, Ortiz de Apodaca Y, Ruiz A. Ascorbic acid in the plasma and blood cells of women with breast cancer. The effect of consumption of food with an elevated content of this vitamin. Nutr Hosp 1995; 10:68–72.Google Scholar
  13. 13.
    Kurbacher CM, et al. Ascorbic acid (vitamin C) improves the antineoplastic activity doxorubicin, cisplatin and paclitaxel in human breast carcinoma cells in vitro. Cancer Lett 1996; 103(2):183–189.PubMedCrossRefGoogle Scholar
  14. 14.
    Valcic S, et al. Inhibitory effects of six green tea catechins and caffeine on the growth of four selected human tumor cell lines. Anticancer Drugs 1996; 7:461–468.PubMedCrossRefGoogle Scholar
  15. 15.
    Mukhtar H, Ahmed N. Tea polypheonols: prevention of cancer and optimizing health. Am J Clin Nutr 2000; 71: 1698S-1720S.PubMedGoogle Scholar
  16. 16.
    Yang GY, Liao J, Kim K, Yurkow EJ, Yang CS. Inhibition of growth and induction of apoptosis in human cancer cell lines by tea polyphenols. Carcinogenesis 1998; 19:611–616.PubMedCrossRefGoogle Scholar
  17. 17.
    Sartippour MR, et al. Green tea and its catechins inhibit breast cancer xenografts. Nutr Cancer 2001; 40:149–156.PubMedCrossRefGoogle Scholar
  18. 18.
    Nakachi K, et al. Influence of drinking green tea on breast cancer malignancy among Japanese patients. Jpn J Cancer Res 1998; 8:254–261.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc 2005

Authors and Affiliations

  • M. Waheed Roomi
    • 1
  • Vadim Ivanov
    • 1
  • Tatiana Kalinovsky
    • 1
  • Aleksandra Niedzwiecki
    • 1
  • Matthias Rath
    • 1
  1. 1.Matthias Rath ResearchCancer Research DivisionSanta ClaraUSA

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