Background
The human T-cell lymphotropic virus type 1 (HTLV1) is a retrovirus that infects CD4-positive T-cells resulting in the development of adult T-cell leukemia (ATL) in approximately 5% of the cases. ATL manifests as a consequence of the clonal expansion of mature and activated infected CD-4 positive T-cells and is associated with a poor prognosis due to its immunosuppressive and chemotherapy-resistant nature [
1]. The causative agent of this disease, HTLV1, is transmitted via breast feeding, transplacentally from mother to child, through sexual contact and blood transfusion [
2]; while in vivo transmission occurs through cell-to-cell contact [
3]. The HTLV1 oncoprotein, Tax, has been shown to be vital for viral persistence and leukemogenesis due to its pleitoropic effects on cellular proliferation and apoptosis as well as viral replication [
4]. As such, targeting Tax has become a novel approach in the treatment of ATL; however, the current options either failed to nullify the issue of relapse [
5]. Therefore, it is essential to unravel a nontoxic compound having an inherent ability to inhibit Tax expression and a potential to serve as a therapeutic agent against this aggressive malignancy.
The successful documentation of the safety and effectiveness of botanical and dietary natural compounds in cancer prevention has led to the development of a nutrient mixture composed of ascorbic acid (AA), lysine, proline, arginine, epigallocatechingallate (EGCG) and other micronutrients [
6]. This natural assortment of nutrients, also known as SNS has exhibited synergistic anticancer properties in a large number of solid cancer cell lines, blocking tumor growth, tumor invasion and MMP expression, both in vitro and in vivo [
7]. Not only that, but SNS had an anti-proliferative effect against HTVL-1 positive and negative malignant T-lymphocytes and demonstrated pro-apoptotic effects with respect to HTLV-1 leukemic cells in specific, via the up-regulation of the pro-apoptotic proteins p53, p21 and Bax, and the down-regulation of the pro-survival Bcl2-α protein [
8]. We have also reported enhanced antiproliferative activity of SNS in the presence of polyethylene glycol gold plated nano-particles [
9].
SNS was in fact formulated based on the capacity of the individual components to alter key physiological pathways involved in cancer progression and metastasis [
10]. In fact, ingredients of SNS were reported to inhibit the destruction of the extracellular matrix (ECM), which is a pre-requisite for cancer cell invasion and metastasis [
11]. For example, the biosynthesis of collagen depends on an adequate supply of AA, the amino acids lysine and proline as well as the micronutrients manganese and copper [
12]. Therefore, the integration of these nutrients into the formulation would result in strengthening the ECM. Not only that, but lysine is also a natural inhibitor of plasmin-induced proteolysis and, therefore, increases ECM stability by inhibiting the breakdown of collagen fibers [
11]. Similarly, N-acetyl cysteine, AA, selenium and EGCG inhibited the invasiveness of tumor cells by blocking the activity of MMPs, which are a unique family of more than 20 proteases responsible for the proteolytic degradation of the ECM, which is essential for the dissemination of cancer cells to secondary sites [
12]. The over-expression of MMPs, which is a common occurrence in malignant tumors, is correlated with tumor aggressiveness, stage and prognosis [
11,
12].
Additionally, multiple studies have associated the makeup of SNS with inhibitory activity against the transcription factor NF- κB, which is constitutively activated by HTLV-1 Tax protein and has as a critical role in the pathophysiology of ATL [
13]. In fact, the inhibitory effect of EGCG on the activity of NF-kB has recently been demonstrated in ATL cell lines [
14]. With respect to AA, it was able to dose-dependently repress activation of NFκB [Harakeh et al., unpublished data]. Furthermore, N-acetyl-cysteine (NAC) was found to reverse NF-κB binding to DNA and NF-κB-dependent oncogene expression in HaCaT cells [
15]; while selenium compounds were capable of preventing esophageal carcinogenesis by inhibiting NF-kB activation [
16]. Finally, magnesium deficiency induced NFκB expression in endothelial [
17] and macrophage cells [
18]. Nonetheless, the synergistic effect of the nutrient mixture on NF-kB activity in HTLV1 infected cells is yet to be determined.
The combination of these micronutrients is accompanied by the advantage of using lower doses of the separate constituents and has the potential of increasing the biological effect by establishing novel metabolic targets [
6]. Therefore 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 in two HTLV-1 positive cell lines, HuT-102 and C91-PL thus determining the potential of using SNS in conjunction to ongoing treatment.
Acknowledgement
Authors would like to thank Dr. A. Gessain, Institut Pasteur Paris, France for kindly providing the cell lines. In addition, authors would like to thank the American University of Beirut, Biology Department, Lebanon for providing the research fund and King Fahd Medical Research Centre, King Abdulaziz University, Saudi Arabia for technical support. The article is an expansion of an abstract presented in the 2nd IGMC, Nov 2013, Jeddah, KSA.
Declarations
Publication charges for this article have been funded by the Center of Excellence in Genomic Medicine Research (CEGMR), King Abdulaziz University, Jeddah, 21589, Kingdom of Saudi Arabia.
Competing interest
All authors declare no competing interests
Authors’ contributions
SH designed and supervised the study. AN and MR involved in study design. RA, EB and MDA performed the experiments and executed the results. SH, RA, MA and MAE contributed in data analysis and manuscript drafting and editing. EA, GAD and TK participated in data analysis and critically revised the manuscript. All authors have read and approved the final version of the manuscript.