Synergistic Antitumor Effect of Ascorbic Acid, Lysine, Proline, and Epigallocatechin Gallate on Human Fibrosarcoma Cells HT-1080

M.W. Roomi, V.Ivanov, T.Kalinovsky, A. Niedzwiecki, M.Rath
Annals of Cancer Research and Therapy 2004, 12(1-2): 146-156

Matrix metalloproteinases (MMPs) have received much attention in recent years for their role in various malignancies, and have been implicated in tumor invasion, metastasis and angiogenesis. Certain MMPs, such as MMP-2 and MMP-9, have shown a special propensity for promoting cancer. Development of MMP inhibitors has been a recent approach to controlling cancer and blocking metastasis.

We investigated the effect of EGCG individually and in combination with lysine, proline, and ascorbic acid (LPA), in vitro, on human fibrosarcoma cells HT-1080, by measuring: cytotoxicity, modulation of MMP-2 and MMP-9, and invasive potential. Fibrosarcoma cell line was chosen for this study as it expresses both MMP-2 and MMP-9. Cytotoxicity was evaluated based on cell proliferation by MTT assay and MMP expression in condition media by gelatinase zymography. Invasion was evaluated through Matrigel. Treatment of fibrosarcoma cells with EGCG independently showed dose-dependent cytotoxicity that was enhanced when EGCG was combined with LPA, with a maximum toxic effect of 45% at 50 µg/ml EGCG + LPA. Zymography showed dose dependent inhibition of MMP-2 and MMP-9 expression by EGCG, also enhanced at each concentration when combined with LPA, with virtual total inhibition of MMP-2 at LPA + EGCG 20µg/ml and MMP-9 at LPA + EGCG 50 µg/ml concentration. The invasion of fibrosarcoma cells through Matrigel was significantly reduced (63%) with LPA + EGCG 20 µg/ml and totally inhibited with LPA + EGCG 50 µg/ml.Our results suggest that the synergistic effect of lysine, proline, and EGCG, is an effective, yet safe agent for adjunctive therapeutic use in the treatment of fibrosarcoma, by inhibiting cell proliferation, MMP expression, and matrigel invasion.

Key Words:
Antitumor activity, ascorbic acid, cell proliferation, cytotoxicity, EGCG, fibrosarcoma, lysine, Matrigel, invasion, matrix metalloproteinases, MMP, MMP-2, MMP-9, MMP-inhibitors, MTT assay, nutrient synergy, proline, toxicity

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