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In vitro anticarcinogenic effect of a nutrient mixture on human rhabdomyosarcoma cells

M.W. Roomi, V. Ivanov, T. Kalinovsky, A. Niedzwiecki, M. Rath
Gene Therapy and Molecular Biology 2007, 11: 133-142

Rhabdomyosarcoma, the most common pediatric soft tissue sarcoma of mesenchymal origin, has metastasized in ~25% of all patients at time of diagnosis. Though current treatment strategies have achieved some success, they are associated with severe adverse effects. We investigated the effect of a nutrient mixture (NM), which has shown antitumor effects on various cancer cell lines, on rhabdomyosarcoma cell growth, apoptosis, MMP secretion, and invasion. Human rhabdomyosarcoma cells, grown in DME, were treated at near confluence with NM at 0, 10, 50, 100, 500 and 1000 μg/ml in triplicate at each dose.

MMP secretion was studied by zymography, viability by MTT assay, cell invasion through Matrigel, and morphology and apoptosis by H&E staining and live green caspase kit. Zymography demonstrated MMP-2 secretion and PMA-induced MMP-9 secretion. NM inhibited the secretion of both MMPs in a dose-dependent fashion, with virtual total inhibition at 500 μg/ml NM. Cell invasion through Matrigel was inhibited at 10, 50, 100 and 500 μg/ml by 75%, 80%, 92% and 100% (p = 0.02) respectively. NM was slightly toxic at 1000 μg/ml (20% over control, p = 0.016) to rhabdomyosarcoma cells. Cells exposed to NM showed dose-dependent apoptosis with 90% of cells in late apoptosis at 1000 μg/ml. These results suggest that NM has potential in the treatment of rhabdomyosarcoma by inducing cell apoptosis and inhibiting cell invasion and MMP secretion without toxic effects.

Key Words:
Rhabdomyosarcoma, apoptosis, MMPs, Matrigel invasion, nutrients, green tea extract, ascorbic acid, lysine

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