Effects of Ascorbic Acid on Tax, NF-κB and MMP-9 in Human T-cell Lymphotropic Virus Type 1 Positive Malignant T-Lymphocytes

Authors: Steve Harakeh,*, Jihane Khalife, Elias Baydoun, Rania Azar, Ahmed Al- Hejin, Elie Barbour,
Esam Azhar, Aleksandra Niedzwiecki, Soad Al Jaouni, Mona Diab-Assaf, and Mathias Rath

Journal Name: Anti-Cancer Agents in Medicinal Chemistry, 2017, 17, 000-000

DOI: 10.2174/1871520617666170725160628

Abstract: 

HTLV-1 is a retrovirus that infects CD4-positive cells and leads to Adult T-cell leukemia by constitutive activation of nuclear factor kappa B. Ascorbic acid (AA) is an essential nutrient that possess anti-proliferative and pro-apoptotic activity against a number of malignant cell lines. This study delineates the effect of AA on Tax protein expression as well as NF-κB and MMP9 activity in two HTLV-1-positive leukemia cells (HuT-102 and C91-PL).

Methods: The cytotoxic and antiproliferative effect of AA were studied by LDH release and MTT tests, respectively. The proteins expression level was assessed by western blotting. RT-PCR was used to study mRNAs level. Finally, ELISA/EMSA and Zymography were used to evaluate NF-κB and MMP-9 activities, respectively.

Results: Cell lines were treated with non-cytotoxic concentrations of AA for 48h and 96h, which resulted in a significant inhibition of proliferation at a concentration of 50μg/ml at 96h in both cell lines. The same concentration inhibited Tax protein expression as well as the NF-κB nuclearization and DNA binding activity. The inhibitory effect of AA on MMP9 protein expression and activity started at 100μg/ml and 50μg/ml in HuT-102 and C91-PL cells respectively, with no effect at the transcriptional levels of MMP-9 in either one of the two cell lines.

Conclusion: These results indicated that while AA exerted its anti-proliferative effect on the NF- κB activation pathway by suppressing Tax expression, its effects on MMP9 seemed to be independent of this mechanism and follow a different approach.


Key words: Acute T-cell leukemia, Human T-cell Lymphotrophic Virus type I (HTLV-1), Tax, NF-κB pathway, Matrix metalloproteinase-9 (MMP-9), Ascorbic Acid (AA).
Access:

 

 

Effect Of A Nutrient Mixture On Fanconi Anemia Fibroblast And Normal Human Dermal Fibroblast: A Comparison

M.W. Roomi, T. Kalinovsky, A. Niedzwiecki and M. Rath
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050
Open Journal of Apoptosis 5:2-8, 2016

Abstract: 

Fanconi anemia (FA) is a fatal heterogeneous autosomal recessive disorder, characterized by progressive bone marrow failure, congenital defect and cancer predisposition. Cell culture from FA fibroblast (FAF) displays certain abnormalities as compared to normal human dermal fibroblast (NHDF). This prompted us to investigate the effect of a novel nutrient mixture (NM) containing ascorbic acid, lysine, proline and green tea extract, which has demonstrated a broad spectrum of pharmacological activities, on FAF compared to NHDF.

Read more ...

In vitro inhibition of matrix metalloproteinases, invasion and growth of Fanconi anemia human FANCA and FANCC lymphoblasts by nutrient mixture

Roomi MW,Bhanap B, Roomi NW, Niedzwiecki A, Rath M.

Exp Oncol.2014;36(3):212-4.

Abstract: 

Aim: Fanconi anemia is a rare genetic disorder with high propensity for development of cancers, such as aplastic anemia, leukemia and head and neck cancers. Collagen digesting matrix metalloproteinase (MMP) enzymes have been implicated in for their role in various malignancies and to promote metastasis.

Read more ...

Epigallocatechin-3-gallate Inhibits Tax-dependent Activation of Nuclear Factor Kappa B and of Matrix Metalloproteinase 9 in Human T-cell Lymphotropic Virus-1 Positive Leukemia Cells.

Steve Harakeh, Mona Diab-Assaf, Rania Azar, Hani Mutlak Abdulla Hassan, Safwan Tayeb, Khalil Abou-El-Ardat, Ghazi Abdullah Damanhouri, Ishtiaq Qadri, Adel Abuzenadah, Adeel Chaudhary, Taha Kumosani, Aleksandra Niedzwiecki, Mathias Rath, Haitham Yacoub, Esam Azhar, Elie Barbour
Asian Pacific journal of cancer prevention: APJCP (Impact Factor: 1.27). 01/2014; 15(3):1219-1225.

Abstract:
Epigallocatechin-3-gallate (EGCG) is the most abundant polyphenol molecule from green tea and is known to exhibit antioxidative as well as tumor suppressing activity. In order to examine EGCG tumor invasion and suppressing activity against adult T-cell leukemia (ATL), two HTLV-1 positive leukemia cells (HuT-102 and C91- PL) were treated with non-cytotoxic concentrations of EGCG for 2 and 4 days.

Read more ...

Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human Fanconi anemia fibroblast cell lines

Roomi MW,Bhanap B, Roomi NW, Niedzwiecki A, Rath M.

ExpOncol. 2013; 35(1):20-24.

Abstract:

Aim: Fanconi Anemia, an autosomal recessive disorder, is characterized by chromosomal abnormality leading to birth defects, progressive bone marrow failure, and a high probability of developing malignancy at an early age. Head and neck squamous cell carcinoma and myeloid leukemia are the major causes of cancer related morbidity and mortality in Fanconi anemia patients.

Read more ...

Inhibition of growth and expression of inflammation mediators in human leukemic cell line u-937 by a nutrient mixture

M.W. Roomi, T. Kalinovsky, N.W. Roomi, M. Rath and A. Niedzwiecki
Dr. Rath Research Institute, Cancer Division, Santa Clara, CA, USA
Experimental Oncology 2013; 35(3): 180-186

Abstract:
A nutrient mixture (NM) containing ascorbic acid, lysine, proline and green tea extract has exhibited anticancer activity in vitro and in vivo in a number of cancer cell lines. We investigated the effect of NM on human leukemic myeloid U-937 cells in vitro by measuring: cell proliferation, MMP expression, invasion, apoptosis, and COX-2 and COX-1 protein expression. Human leukemic cell line U-937 (ATCC) was cultured in RPMI medium supplemented with fetal bovine serum and antibiotics.

Read more ...

In vitro and in vivo inhibition of human Fanconi anemia head and neck squamous carcinoma by a novel nutrient mixture

M.W. Roomi, T. Kalinovsky, N.W. Roomi, A. Niedzwiecki and M. Rath
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050
International Journal of Oncology 2012; 41(6), 1996-2004

Abstract
Head and neck squamous cell carcinoma (HNSCC) and acute myeloid leukemia are the major causes of mortality and morbidity in Fanconi anemia (FA) patients. The objective of this study was to investigate the antineoplastic activity of NM, a novel antineoplastic nutrient mixture (containing lysine, proline, ascorbic acid and green tea extract) on human FA HNSCC in vitro and in vivo. Human FA HNSCC cell line OHSU-974 (Fanconi Anemia Research Fund) was cultured in RPMI medium supplemented with 20% FBS and antibiotics. At near confluence, cells were treated in triplicate with different concentrations of NM: 0, 50, 100, 250, 500 and 1000 µg/ml.

Read more ...

In vivo and in vitro antitumor effects of nutrient mixture in murine leukemia cell line P-388

Roomi MW, Roomi NW, Bhanap B, Rath M, Niedzwiecki A                                                        

ExpOncol. 2011; 33(2):71-7

Abstract:

Aim: Leukemia is characterized by uncontrolled marrow cell proliferation and metastatic foci. We investigated the antitumor potential of a nutrient mixture on malignant leukemia P-388 cells. Methods: The nutrient mixture containing lysine, proline, ascorbic acid, green tea extract and other nutrients is formulated to target key pathways in cancer progression.

Read more ...

Nutrient Mixture Inhibits in vitro and in vivo Growth of Human Acute Promyelocytic Leukemia HL-60 Cells

MW Roomi, NW Roomi, B Bhanap, A Niedzwiecki, M Rath;                                                        

Exp Oncol. 2011;33(4):212-215

Abstract:

Aim:Untreated acute promyelocytic leukemia is the most malignant form of acute leukemias, with median survival of less than one month. We investigated in vitro and in vivo synergistic effects of a nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea extract, on acute promyelocytic leukemia HL-60 cells.

Read more ...

Antineoplastic effects of nutrient mixture on raji and jurkat T cells: the two highly aggressive Non-Hodgkin's Lymphoma cell lines

Roomi MW, Bhananp BA, Roomi NW, Rath M, Niedzwiecki A
Experimental Oncology 2009; 31(3): 149-155

Abstract:
Non-Hodgkin lymphomas incidence has increased more than 70% in last 25 years. Aggressiveness, higher relapse rate, and treatment complications pose significant barriers. Decreased food intake and side effects of treatments make cancer patients vulnerable to deficiency of essential nutrients such as vitamin C, lysine, and proline leading to the formation of weak extra cellular matrix susceptible to easy breakdown by matrix metalloproteinase enzymes.

Read more ...

Antineoplastic effect of nutrient mixture on Raji and Jurkat T cells: the two highly aggressive non Hodgkin’s lymphoma cell lines

MW Roomi, BA Bhanap, NW Roomi, A. Niedzwiecki and M. Rath

Experimental Oncology 2009; 31(3): 149-155.

Abstract:

Non-Hodgkin lymphomas incidence has increased more than 70% in last 25 years. Aggressiveness, higher relapse rate, and treatment complications pose significant barriers. Decreased food intake and side effects of treatments make cancer patients vulnerable to deficiency of essential nutrients such as vitamin C, lysine, and proline leading to the formation of weak extra cellular matrix susceptible to easy breakdown by matrix metalloproteinase enzymes. Inhibition of these enzymes has shown promise in stopping metastasis.

Read more ...

Epigallocatechin-3-gallate induces apoptosis and cell cycle arrest in HTLV-1-positive and -negative leukemia cells

Harakeh S, Abu-El-Ardat K, Diab-Assaf M, Niedzwiecki A, El-Sabban M, Rath M 

Medical Oncology 2008; 25 (1): 30-39

Abstract:
The objective of this study is to evaluate the efficacy of epigallocatechin gallate against ATL cells. The anti-proliferative and pro-apoptotic effects of EGCG were evaluated in HTLV-1-positive and -negative cells. EGCG exhibited a marked decrease in proliferation of ATL cells at 96 h of treatment.The results indicated that TGF-alpha was down regulated whereas levels of TGF-beta2 increased. Cell cycle distribution analysis revealed an increase in cells in the pre-G(1) phase which was confirmed by ELISA. The results on proteins showed an up-regulation of p53, Bax and p21 protein levels while the levels of Bcl-2alpha were down regulated.

The full study is available online at:

http://www.springerlink.com/content/63368t4025p27312

Ascorbic acid induces apoptosis in adult T-cell leukemia

S. Harakeh, M. Diab-Assaf, J.C. Khalife, K.A. Abu-el-Ardat, E. Baydoun, A. Niedzwiecki, M.E. El-Sabban, M. Rath
Anticancer Research 2007; 27: 289-298

Adult T-cell leukemia (ATL) is an acute malignancy of activated T-cells caused by the human T-cell lymphotrophic virus type-1 (HTLV-1). The effects of non-cytotoxic concentrations of ascorbic acid (AA) were evaluated against HTLV-1 positive and negative cells. The effect of AA on apoptosis and proliferation was evaluated by cell cycle analysis.

Read more ...

Mechanistic aspects of apoptosis induction by L-lysine in both HTLV-1-positive and -negative cell lines

S. Harakeh, M. Diab-Assaf, K. Abu-El-Ardat, A. Niedzwiecki, M. Rath
Chemico-Biological Interactions 2006, 164: 102-114

Abstract:
The retrovirus human T-cell lymphotrophic virus type-1 (HTLV-1) causes adult T-cell leukemia (ATL), which remains with no cure. This study evaluates the effects of L-lysine on proliferation and induction of apoptosis using non-cytotoxic concentrations of the test compound against HTLV-1 positive and negative malignant cell lines.

Read more ...

Apoptosis induction by Nurient 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 2006, 30: 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.

Read more ...