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In Vivo and In Vitro Effect of a Nutrient Mixture on Human Hepatoma Cell Line SK‐Hep‐1
Author(s) -
Roomi M. Waheed,
Ivanov Vadim,
Niedzwiecki Aleksandra,
Rath Matthias
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a757-d
Subject(s) - in vivo , zymography , in vitro , apoptosis , ascorbic acid , chemistry , microbiology and biotechnology , matrigel , cell culture , pharmacology , biology , matrix metalloproteinase , biochemistry , genetics , food science
Worldwide, hepatocellular carcinoma (HCC) is the sixth most common cancer and third most common cause of death from cancer. There is no effective treatment for liver cancer, and the survival rates are low with only 6% in the U.S. surviving five years. A novel nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract has exhibited anti‐tumor activity in vivo and in vitro. We investigated the effect of NM on human hepatoma cell line in vitro and in vivo. After one week of isolation, 5–6 weeks old male athymic nude mice were inoculated with 3×10 6 SK‐Hep‐1 cells subcutaneously and randomly divided into two groups; group A was fed a regular diet and group B a regular diet supplemented with 0.5% NM. 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 SK‐Hep‐1 cells, measuring cell proliferation by MTT assay, invasion through Matrigel, apoptosis by green caspase detection kit, MMP secretion by zymography, and morphology by H&E staining. NM inhibited the growth of SK‐Hep‐1 xenograft tumors by 50%. In vitro, NM exhibited 30% toxicity over the control at 500 and 1000 μg/ml concentration. Zymography demonstrated MMP‐2 and MMP‐9. NM inhibited the secretion of both MMPs in a dose dependent fashion, with virtual total inhibition at 1000 μg/ml. Invasion was inhibited at 10, 50, 100, 500 and 1000 μg/ml by 40%, 46%, 55%, 84% and 100% respectively. NM induced slight apoptosis at 100, considerable at 500 and significant at 1000 μg/ml NM. H&E staining showed no morphological changes up to 100 μg/ml. These results suggest that NM has therapeutic potential in treatment of HCC.