Dietary restriction of two amino acids, tyrosine (Tyr) and phenylalanine (Phe), suppresses the malignant phenotype of B16-BL6 (BL6) murine melanoma. Our data indicate that Tyr and Phe restriction inhibits the ability of tumor cells to establish metastatic tumor foci. This proposal focuses on the mechanism responsible for this Tyr and Phe modulation and will test the hypothesis that it is multifactorial, encompassing decreased invasiveness, angiogenesis, and growth response.
The specific aims are to 1) examine invasion of BL6 murine melanoma and Tyr- and Phe-modulated BL6 variants through reconstituted basement membrane and selected matrix components and the response to chemotaxis, 2) examine differential proteolytic enzyme and protease inhibitor activities in media conditioned by BL6 and the modulated variants, 3) examine soluble angiogenic factors released into conditioned media by these tumor cells, and 4) determine if phenotypic modulation by Tyr and Phe restriction is associated with change in cell cycle characteristics, growth factors, or tumor suppressor gene expression. To confirm preliminary results indicating that Tyr and Phe limitation alters invasive characteristics of tumor cells, experiments are designed to specifically examine invasion of cells through substrates of extracellular matrix extracted from lung, Matrigel, collagen types I and IV, laminin, fibronectin, and elastin. To analyze differential release by our Tyr- and Phe-modulated variants of soluble proteolytic enzymes, conditioned media from cell cultures will be tested for ability to degrade collagen types I and IV, laminin, and elastin, in the presence and absence of protease inhibitors. These assays will be extended to determine release and activity in conditioned media of other soluble proteinases which are known to be involved in invasion, specifically cathepsins B and L, plasminogen activator, and chymotrypsin. The possibility of increased production and release of proteinase inhibitors will also be studied. Production and release of angiogenic factors such as fibroblast growth factor, transforming growth factor beta (TGF-beta), and angiogenin by BL6 and Tyr- and Phe-modulated BL6 variants will also be determined. Altered growth patterns and/or growth factor and tumor suppressor gene expression may contribute to the antimetastatic effects of Tyr and Phe restriction. We will investigate whether changes occur in the cell cycle and in expression of TGF-beta and the tumor suppressor genes, nm23 and TIMP (tissue inhibitors of metalloproteinases).

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA042465-06
Application #
3183836
Study Section
Metabolic Pathology Study Section (MEP)
Project Start
1987-07-01
Project End
1996-04-30
Budget Start
1993-05-01
Budget End
1994-04-30
Support Year
6
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Washington State University
Department
Type
Schools of Pharmacy
DUNS #
041485301
City
Pullman
State
WA
Country
United States
Zip Code
99164
Fu, Y M; Yu, Z X; Pelayo, B A et al. (1999) Focal adhesion kinase-dependent apoptosis of melanoma induced by tyrosine and phenylalanine deficiency. Cancer Res 59:758-65
Pelayo, B A; Fu, Y M; Meadows, G G (1999) Inhibition of B16BL6 melanoma invasion by tyrosine and phenylalanine deprivation is associated with decreased secretion of plasminogen activators and increased plasminogen activator inhibitors. Clin Exp Metastasis 17:841-8
Fu, Y M; Li, Y Q; Meadows, G G (1998) Influence of tyrosine and phenylalanine limitation of cytotoxicity of chimeric TGF-alpha toxins on B16BL6 murine melanoma in vitro. Nutr Cancer 31:1-7
Fu, Y M; Yu, Z X; Ferrans, V J et al. (1997) Tyrosine and phenylalanine restriction induces G0/G1 cell cycle arrest in murine melanoma in vitro and in vivo. Nutr Cancer 29:104-13
Elstad, C A; Thrall, B D; Raha, G et al. (1996) Tyrosine and phenylalanine restriction sensitizes adriamycin-resistant P388 leukemia cells to adriamycin. Nutr Cancer 25:47-60
Uhlenkott, C E; Huijzer, J C; Cardeiro, D J et al. (1996) Attachment, invasion, chemotaxis, and proteinase expression of B16-BL6 melanoma cells exhibiting a low metastatic phenotype after exposure to dietary restriction of tyrosine and phenylalanine. Clin Exp Metastasis 14:125-37
Huijzer, J C; McFarland, M; Niles, R M et al. (1996) Phorbol 12-myristate 13-acetate enhances nm23 gene expression in murine melanocytes but not in syngeneic B16-BL6 melanoma variants. J Cell Physiol 166:487-94
Huijzer, J C; Uhlenkott, C E; Meadows, G G (1995) Differences in expression of metalloproteinases and plasminogen activators in murine melanocytes and B16 melanoma variants: lack of association with in vitro invasion. Int J Cancer 63:92-9
Elstad, C A; Meadows, G G; Aslakson, C J et al. (1994) Evidence for nutrient modulation of tumor phenotype: impact of tyrosine and phenylalanine restriction. Adv Exp Med Biol 354:171-83
Blanckaert, V D; Schelling, M E; Elstad, C A et al. (1993) Differential growth factor production, secretion, and response by high and low metastatic variants of B16BL6 melanoma. Cancer Res 53:4075-81

Showing the most recent 10 out of 18 publications