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A Natural Anti-Cancer Formula - A Specific Formulation of Nutrients Containing Lysine,Proline, Ascorbic Acid, and Epigallocatechin Gallate Inhibits Matrix Metalloproteinases Activity and Invasion Potential of Human Cancer Cell Lines.

M.W. Roomi, S.P. Netke, V. Ivanov, M. Rath and A. Niedzwiecki

Presented at: European Organization for Research and Treatment of Cancer (EORTC), AACR and NCI Symposium on Molecular Targets and Cancer Therapeutics, Frankfurt, Germany, Nov 19-22, 2002
Published in: European Cancer Journal, 38, Suppl.7/Abs.280, 2002

One of the hallmarks of cancer is its ability to invade and metastasize to distal organs. Matrix metalloproteinases ( MMPs ) have been identified as key players in tumor invasion and metastasis. Excessive MMPs secretion has been regarded as an index of malignancy which leads to the degradation of extra cellular matrix. Current treatment protocols with chemotherapy and/or radiation although beneficial, are toxic and have the potential to destroy healthy cells as well. Our approach has been to develop strategies to inhibit cancer development, progression and metastasis using naturally occurring nutrients, which are relatively non-toxic. Lysine and proline are building blocks of collagen fibers that stabilize connective tissue by inhibiting the enzymatic digestion of collagen fibers. Vitamin C is essential for production of collagen and acts as a powerful antioxidant by scavenging free radicals and thereby protects cells from damage. Epigallocatechin gallate (EGCG), is a green tea extract with antioxidant and anticancerogenic properties. It prevent cancer cell invasion by inhibiting MMPs. It is postulated that the combination of these nutrients would exert a very potent synergistic anticancer activity.

Based on the above prediction, a natural anti-cancer formula containing a mixture of nutrients such as lysine, proline, ascorbic acid and EGCG was formulated. In the present study, we investigated the effect of a natural anti-cancer formula on MMPs expression, matrix invasion potential and cell proliferation in several human cancer lines those of skin (melanoma), breast (MDA MB 231) and liver (Hep G2). We also studied the effects of a natural anti-cancer formula on normal human dermal fibroblast (NHDF) and on co-culture of melanoma and NHDF cells. MMPs expression was studied by zymography, extracellular matrix invasion by using reconstituted basement membrane (Matrigel ) and cytotoxicity/cell proliferation by MTT assay. A natural anti-cancer formula inhibits the expression of both MMP-2 and MMP-9 in a dose dependent fashion. The expression of both MMP -2 and –9 were significantly inhibited with a concentration of 100 µg/ml of a natural anti-cancer formula and virtually not detectable with a concentration of 1000 µg/ml. A natural anti-cancer formula used at 10 and 100 µg/ml concentrations did not significantly affect cells viability and at 1000 µg/ml it showed cytotoxicity at the range of 10-40% depending on the cell type.

The invasion of melanoma cells through Matrigel matrices was inhibited by 20% and 100% at 10 and 50 µg/ml respectively. Similar invasion of MDA MB 231 was also reduced by 50%, 60% and 95% at 10, 50 and 100 µg/ml respectively. When melanoma cells were co-cultured with NHDF cells, a natural anti-cancer formula inhibited the invasion by 30% and 100% with 10 and 50 µg/ml respectively.

Thus these results demonstrate that a natural anti-cancer formula is very effective for several cancer cell lines and also in co-culture in inhibiting the expression of MMPs and preventing cellular invasion through Matrigel. These observations revealed that a natural anti-cancer formula may provide a natural therapeutic basis which makes it a valuable and promising candidate for the treatment of human cancers. Currently, experiments are in progress to evaluate the efficacy of a natural anti-cancer formula in a clinical setting.



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