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Anticancer Activity of a Nutrient Mixture inMalignant Leukemia Cell Line P-388 in vitro#96

M.W. Roomi, V. Ivanov, A. Niedzwiecki and M. RathMatthias Rath Research Institute, Cancer Division, Santa Clara, CA 95050

Presented at: International Society for Biological Therapy of Cancer, Nov 10-13,2005, Alexandria, VA

Published in: International Society for Biological Therapy of CancerProceedings, Abstract #96

Abstract

Introduction:A unique nutrient formulation (NM) consisting of lysine, proline, ascorbic acid andgreen tea extract has been shown to exhibit anti-tumor effects in several cancerlines both in vitro and in vivo studies. NM acts by exerting a potent synergisticanticarcinogenic effect by inhibiting proliferation, MMP expression, and invasionthrough Matrigel. Leukemia, acute or chronic neoplasia, is a disorder of bonemarrow in which unrestrained proliferation of white blood cells occurs, resultingin formation of metastatic colonies and penetration through matrix barriers andblood vessel walls, a process mediated through MMPs.

Objective:In the present investigation we examined the anticancer effect of NM on murineleukemia cell line P-388 on proliferation, MMP expression and invasion throughMatrigel

Materials and Methods:Murine leukemia cell line P-388D1 (ATCC) was grown in DEM media in 10% horseserum and antibiotics, and treated with NM at 0, 10, 50, 100, 500, and 1000g/ml concentration in triplicate at each dose. Cell proliferation was assessed byMTT assay, invasion through Matrigel, and MMPs by gelatinase zymography. Cells

were also treated with PMA (200 ng/ml) for MMP-9 induction.

Results:NM exhibited slight toxicity at 50 g/ml and significant (70% and 90% over thecontrol) antiproliferative effect at 500 and 1000 g/ml respectively,. Invasionthrough Matrigel was inhibited at 500 and 1000 g/ml by 80% and 100%respectively. However, no MMP (-2 or 9) activity was detected even with PMAtreatment of P-388 cells.

Conclusion:Our results suggest that NM is a promising new therapeutic agent for leukemia,

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and is a potential candidate for human trials.

Comment:Leukemia, a disorder of bone marrow characterized by unrestrained whiteblood cell proliferation, results in formation of metastatic colonies andpenetration through matrix barriers and blood vessel walls. Degradationand invasion through matrix barriers and vessel walls is mediated bysecretion of MMPs. A nutrient mixture (NM) containing lysine, proline,ascorbic acid and green tea extract exhibited significant inhibition of

Matrigel invasion (with total block at 1000 g/ml) and significantantiproliferative effect at 500 g/ml (70%) and 1000 g/ml (90%). Theseresults are important as they indicate NM is a promising therapeutic agentfor leukemia.

2006 Matthias Rath Research - All Rights Res