title: Ascorbate supplementation inhibits growth and metastasis of B16FO melanoma             and 4T1 breast cancer cells in vitamin C-deficient mice.
abstract: Degradation of the extracellular matrix (ECM) plays a critical role in the             formation of tumors and metastasis and has been found to correlate with the aggressiveness             of tumor growth and invasiveness of cancer. Ascorbic acid, which is known to be             essential for the structural integrity of the intercellular matrix, is not produced             by humans and must be obtained from the diet. Cancer patients have been shown             to have very low reserves of ascorbic acid. Our main objective was to determine             the effect of ascorbate supplementation on metastasis, tumor growth and tumor             immunohistochemistry in mice unable to synthesize ascorbic acid [gulonolactone             oxidase (gulo) knockout (KO)] when challenged with B16FO melanoma or 4T1 breast             cancer cells. Gulo KO female mice 36-38 weeks of age were deprived of or maintained             on ascorbate in food and water for 4 weeks prior to and 2 weeks post intraperitoneal             (IP) injection of 5x105 B16FO murine melanoma cells or to injection of 5x105 4T1             breast cancer cells into the mammary pad of mice. Ascorbate-supplemented gulo KO             mice injected with B16FO melanoma cells demonstrated significant reduction (by             71%, p=0.005) in tumor metastasis compared to gulo KO mice on the control diet.             The mean tumor weight in ascorbate supplemented mice injected with 4T1 cells was             reduced by 28% compared to tumor weight in scorbutic mice. Scorbutic tumors demonstrated             large dark cores, associated with increased necrotic areas and breaches to the             tumor surface, apoptosis and matrix metalloproteinase-9 (MMP-9), and weak, disorganized             or missing collagen I tumor capsule. In contrast, the ascorbate-supplemented group             tumors had smaller fainter colored cores and confined areas of necrosis/apoptosis             with no breaches from the core to the outside of the tumor and a robust collagen I             tumor capsule. In both studies, ascorbate supplementation of gulo KO mice resulted             in profoundly decreased serum inflammatory cytokine interleukin (IL)-6 (99% decrease,             p=0.01 in the B16F0 study and 85% decrease, p=0.08 in the 4T1 study) compared             to the levels in gulo KO mice deprived of ascorbate. In the B16FO study, ascorbate             supplementation of gulo KO mice resulted in profoundly decreased serum VEGF (98%             decrease, p=0.019 than in the scorbutic gulo KO mice). As expected, mean serum             ascorbate level in ascorbate-restricted mice was 2% (p<0.001) of the mean ascorbate             levels in supplemented mice. In conclusion, ascorbate supplementation hinders             metastasis, tumor growth and inflammatory cytokine secretion as well as enhanced             encapsulation of tumors elicited by melanoma and breast cancer cell challenge             in gulo KO mice.
authors.affiliation: Dr Rath Research Institute, Santa Clara, CA 95050, USA.
authors.forename: John
authors.initials: J
authors.lastname: Cha
authors.forename: M Waheed
authors.initials: MW
authors.lastname: Roomi
authors.forename: Vadim
authors.initials: V
authors.lastname: Ivanov
authors.forename: Tatiana
authors.initials: T
authors.lastname: Kalinovsky
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Niedzwiecki
authors.forename: Matthias
authors.initials: M
authors.lastname: Rath
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000008:administration & dosage* / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D001205:Ascorbic Acid / Q000008:administration & dosage* / Q000378:metabolism; D001206:Ascorbic Acid Deficiency / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D049109:Cell Proliferation / Q000187:drug effects; D019587:Dietary Supplements; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D050544:L-Gulonolactone Oxidase / Q000502:physiology*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008546:Melanoma, Experimental / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D009362:Neoplasm Metastasis; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D000975:Antioxidants; D042461:Vascular Endothelial Growth Factor A; D050544:L-Gulonolactone Oxidase; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D001205:Ascorbic Acid