M.W. Roomi, N.W. Roomi, M. Rath and A. Niedzwiecki
Dr. Rath Research Institute, Cancer Division, Santa Clara, CA 95050
23rd Annual Fanconi Anemia Research Fund Scientific Symposium, Barcelona, Spain, October 20-23, 2011.
Proceedings of the 23rd Annual Fanconi Anemia Research Fund Scientific Symposium, page 139.
Fanconi Anemia (FA), an autosomal recessive disease associated with chromosomal instability, is characterized by progressive bone marrow failure, birth defects and high propensity to develop malignancies. Head and neck squamous cell carcinoma (HNSCC) and acute myeloid leukemia are the major causes of mortality and morbidity in 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) that demonstrates a broad spectrum of antitumor activity against a number of human cancer cell lines, on human FAHNSCC in vitro and in vivo.
Methods and Materials:
Human FAHNSCC cell line OHSU-974 (Fanconi Anemia Research Fund, Oregon Health & Science University, Portland, Oregon, USA) 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. Cells were also treated with PMA to induce MMP-9 activity. Cell proliferation was detected by MTT assay, secretion of MMPs by gelatinase zymography, invasion through Matrigel and morphology by H&E staining. In vivo, athymic male nude mice (n=12) were inoculated with 3x106 OHSU-974 cells subcutaneously and randomly divided into two groups: group A was fed a regular diet and group B a regular diet supplemented with 1% NM. Four weeks later, the mice were sacrificed and their tumors were excised, weighed and processed for histology.
NM inhibited the growth of OHSU-974 tumor by 48% and tumor burden by 50%. At lower concentrations, NM demonstrated no effect on proliferation, but at 1000 µg/ml showed 40% toxicity. Zymography revealed MMP-2 and PMA-induced MMP-9 secretion. NM suppressed secretion of both MMPs in a dose-dependent manner, with virtual inhibition at 500 µg/ml. NM inhibited OHSU-974 invasion through Matrigel in a dose-dependent fashion with total block at 1000 µg/ml. H&E staining showed no morphological changes below 500 µg/ml.
The results suggest that NM has potential therapeutic use in the treatment of human FA HNSCC.
Current treatment methods for Fanconi Anemia-associated cancers are generally ineffective. Thus, there is a need for development of effective therapeutic agents for these cancers with minimal toxicity. Our studies demonstrated that NM significantly inhibited the growth and tumor burden of FA HNSCC cell line OHSU-974 in vivo. In addition, invasive parameters, such as OHSU-974 cell line MMP-2 and -9 secretion and invasion were significantly inhibited by NM in vitro. These findings suggest potential of NM in treatment of FA HNSCC.