Apoptosis induction by Nutrient Synergy in HTLV-1 positive and negative malignant T-cells

S. Harakeh, M. Diab-Assaf, A. Niedzwiecki, J. Khalife, K. Abu-El-Ardat, M. Rath

Leukemia Research 30, (2006) 869-881.

Introduction:
Human T-cell lymphotropic virus type-1 (HTLV-1) is the causative agent of adult T-cell leukemia, a fatal disease with average survival time of less than one year. A novel nutrient formulation, Nutrient Synergy (NS), which contains lysine, proline, ascorbic acid, and green tea extract, has been shown to be effective in inhibiting cell invasion, proliferation and angiogenesis of various solid cancers.

Methods:
The effects of Nutrient Synergy were evaluated on proliferation (by MTT assay), expression of TGF mRNA (by RT-PCR) and induction of apoptosis (by flow cytometry and ELISA based apoptosis assays) using non-cytotoxic concentrations against HTLV-1 positive (HuT-102 & C91-PL) and negative (CEM & Jurkat) cells.

Results:
NS showed anti-proliferative effect as determined by MTT assay and TGF mRNA protein expression using RT-PCR. NS resulted in the down-regulation of TGF-alpha and an up-regulation in TGF-beta2. NS caused a significant increase in apoptotic cells in the preG(1) phase. These results were confirmed using Cell Death ELISA and Annexin V-FITC. Induction of apoptosis was caused by an up-regulation of p53, p21 and Bax protein levels and a down-regulation of Bcl-2alpha protein expression level.

Conclusion:
The relatively non-toxic Nutrient Synergy demonstrated significant anti-proliferative and pro-apoptotic effects in tested HTLV-1 positive and negative adult T-cell leukemia cell lines, warranting further in vivo investigation.