In Vivo and In Vitro Effect of a Nutrient Mixture of Chemopreventive Agents on Human Hepatocarcinoma Cells

M.W. Roomi, N.W. Roomi, T. Kalinovsky, A. Niedzwiecki and M. Rath
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050

Exp Oncol 2010, 32: 84-91


Purpose: Long-term survival of patients with hepatocellular carcinoma (HCC), a common cancer worldwide, remains poor, due to metastasis and recurrence. We investigated the effect of a novel nutrient mixture (NM) of chemopreventive agents containing ascorbic acid, lysine, proline, and green tea extract on human HCC cell line Sk-Hep-1 in vivo and in vitro.


Methods: Five to six week old male athymic nude mice were inoculated with 3x106 Sk-Hep-1 cells subcutaneously and then randomly divided into two groups. Group A was fed a basal mouse chow diet and Group B the basal diet supplemented with 0.5% NM. Four weeks later, the mice were euthanized and their tumors were excised, weighed and processed for histology. In addition, the effect of NM on the viability of SK-Hep-1 cells, by MTT assay, invasion through Matrigel, MMP secretion by zymography and induction of apoptosis by Live-Green Caspases kit were also determined.

Results: Dietary exposure to 0.5% NM resulted in a decrease in the incidence by 33% as well as inhibition in tumor growth by 36%. In vitro, NM inhibited cell viability by 33% over the control at 500 and 1000 µg/ml concentration and induced apoptosis in a dose-dependent manner. NM inhibited secretion of MMP-2 and MMP-9 as well as invasion through Matrigel in a dose dependent fashion, with virtual total inhibition at 1000 µg/ml.

Conclusions: The results presented demonstrate that the nutrient mixture of chemopreventive agents inhibits the growth of hepatocellular carcinoma cells both in vivo and in vitro and invasion in Matrigel.

hepatocellular carcinoma; SK-Hep-1; nutrient mixture of chemopreventive agents; athymic nude mice; apoptosis; invasion

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