Effects of specific nutrients on tax-dependent activation of NF-κB and MMP-9 in human T-cell lymphotropic virus -1 positive malignant T-lymphocytes
BMC Genomics volume 15, Article number: P72 (2014)
Adult T-cell Leukemia (ATL) is a disease with no known cure so farand it is a resistant to chemotherapy. The virus can be transmitted by exchange of bodily fluids through the placenta and from mother to child. Only 5% of those who are infected develop the disease after a long latency period ranging from 30-50 years [1, 2]. The disease manifests itself as an aggressive proliferation of CD4+ cells with the human T-cell Lymphotropic virus type 1 (HTLV-1) . The leukemogenesis of the virus is mainly attributed to the viral oncoprotein, Tax, that activates the Nuclear Factor kappa B (NF-κB)which in turn stimulates the activity and expression of the matrix metalloproteinase-9 (MMP-9)which is important in angiogenesis. Our previous work has shown that using non-cytotoxic concentrations of a Specific Nutrient Synergy (SNS)mixture resulted in the induction of apoptosis in both HTLV-1 positive and negative malignant T-lymphocytes . The objective of this study is to investigate the efficacy of SNS on Tax expression, NF-κB levels as well as on MMP-9 activity and expression both at the transcriptional and translational levels intwo HTLV-1 positive cell lines, HuT-102 and C91-PL.
Materials and methods
Cell growth, experimental design, source of SNS, preparation and storage of stock solution, were previously described by our group .The effects of non-cytotoxic concentrations of SNS ranging from 0-350 μg/ml were evaluated for their efficacy on proliferation, Tax expression, NF-κBmobility and the activity and expression of MMP-9 at 48h and 96hof incubation.Cytotoxicity of EGCG was assayed using CytoTox 96 Non-radioactive and proliferation was measured using Cell Titer96TM Nonradioactive Cell Proliferation kit( MTT- based assay). Elisa and EMSA were used to assess the effect of SNS on NF-κBmobility. Zymography was used to determine the effects of SNS on the activity and secretion of MMP-9. The expression of MMP-9 was done using RT-PCR at the translational level and Immunoblottingat the transcriptional level.
A significantinhibition of proliferation was seen in both cell lines starting at a concentration of 200μg/ml and in a dose dependent manner. SNS induced a dose dependent decrease in Tax expression (Fig.1),which was paralleled by a down-regulation of the nuclearization of NF-κB (Fig.2). This culminated in the inhibition of the activity of MMP-9 and their expression both at the transcriptional and translational levels (Fig.3).
The role of nutrients in the treatment of disease has been overlooked for a long time. Recently, it has been recognized that nutrients play a crucial role in the outcome of the treatment. The results of this study indicate that a specific nutrient synergy targeted multiple levels pertinent to the progression of ATL. Its activity was mediated through the NF-κB pathway, and hence has the potential to be integrated in the treatment of this disease as a natural, yet potent anticancer agent.
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Harakeh S, Diab-Assaf M, Azar R, Tayeb S, Abou-El-Ardat K, Damanhouri GA, Qadri I, Chaudhary A, Kumosani T, Hassan H, Niedzwiecki A, Rath M, Yacoub H, Barbour E: Epigallocatechin-3-gallate inhibits Tax-dependent activation of Nuclear Factor Kappa B and of Matrix Metalloproteinase 9 in Human T-cell Lymphoropic Virus -1 positive leukemia cells. Asian Pac J Cancer Prev. 2014
Boxus M, Willems L: Mechanisms of HTLV-1 persistence and transformation. Br J Cancer. 2009, 101: 1497-1501.
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Harakeh, S., Diab-Assaf, M., Azar, R. et al. Effects of specific nutrients on tax-dependent activation of NF-κB and MMP-9 in human T-cell lymphotropic virus -1 positive malignant T-lymphocytes. BMC Genomics 15 (Suppl 2), P72 (2014). https://doi.org/10.1186/1471-2164-15-S2-P72