Anti-Metastatic Activity of Nutrient Synergy on Human Colon Cancer Cell Line HCT 116 (2003)

Roomi MW, Roomi NW, Ivanov V, Netke SP, Niedzwiecki A, Rath M.

Presented at: International Research Conference on Food, Nutrition and Cancer
Washington D.C., July 17-18, 2003

Published in: conference proceedings

Abstract

Introduction:
Colon cancer, the second leading cause of cancer death in the United States, causes 55, 000 deaths per year, most commonly in men and women at and over the age of 50 years. Over 90 % of all cancer fatalities result from metastasis to other parts of the body. Matrix metalloproteineases (MMP-2 and MMP-9) have been identified as bulldozers that disintegrate the extracellular matrix (ECM) and assist cancer cell invasion and spread to distal organs. Current treatment with chemotherapy and/or radiation, although beneficial, is toxic and can destroy healthy cells. We have developed strategies to inhibit cancer development, progress, and metastasis using combinations of naturally occurring nutrients, such as lysine, proline, arginine, ascorbic acid, and green tea extract to inhibit enzymatic digestion of collagen fibers and stabilize connective tissue. Furthermore, a mixture of natural compounds often produces a synergistically enhanced therapeutic effect.

Objective:
We postulated that Nutrient Synergy (NS), formulated at optimal levels of these nutrients, would exert very potent anticancer activity, by inhibiting expression of MMPs and cancer cell invasion of human colon cancer cells in culture and in nude mice.

Methods and Materials:
Human colon cancer HCT 116 cells were obtained from ATCC and cultured in modified Eagle media supplemented with 10% fetal bovine serum and antibiotic in 24 well cultured plates. At near confluence, the cells were treated with NS, dissolved in media and tested in triplicate at 0, 10, 50, 100, 500, and 1000 ?g/ml concentration. MMP-9 assay was done by gelatinase zymography in conditioned media. Invasion through Matrigel was assessed. Cell proliferation/cytotoxicity, evaluated by cell number, was measured by MTT assay after 24 hours. Morphology was observed using Hematoxylin and Eosin staining. The effect of NS on tumor growth was studied in five-week old male nude mice injected subcutaneously with 2 X 106 colon cancer cells (HCT 116) in 0.2% PBS + 0.1% matrigel, and fed with a control diet or with a diet containing 0.5% NS. The treated mice were inoculated with 0.5% NS. The mice were sacrificed four weeks after injection, and growth of colon cancer cells was evaluated by tumor weight.

Results:
NS inhibited the expression of MMP-9 in a dose dependent fashion, with significant inhibition at 100 ?g/ml of NS. The invasion of colon cancer cells (HCT 116) through Matrigel was reduced by 54, 66, 76 and 100% with 10, 50, 100 and 500 ?g/ml concentration of NS, respectively. NS was not toxic to colon cancer cells even at 1000 ?g/ml concentration. H&E stains did not show any alterations in morphology with different doses of NS. NS inhibited growth of colon cancer cells by 65% in nude mice.

Conclusion:
In summary, these observations suggest that Nutrient Synergy is a valuable and promising agent with potential antimetastatic activity, by inhibiting invasion, MMP-9 activity, and the growth of colon cancer tumors in nude mice.