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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.W. Roomi, V.Ivanov, T.Kalinovsky, A. Niedzwiecki, M.Rath
Medical Oncology 2005, 22(2): 129-38

Abstract:
Current treatments are generally ineffective once breast cancer has metastasized; median survival is reduced to 2-3 years. 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. 5-6 week old female mice were inoculated with 3x106 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. 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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