Abstract

The proposed study is designed to elucidate the transport and metabolism of methotrexate and the folate coenzymes in hepatic tissue. A monolayer culture system of rat hepatocytes and a rat hepatoma (H35) cell line will be used to examine the transport processes. A comparative assessment will be made between the mormal and transformed cells and the normal cells will be examined to determine the properties that allow these cells to act as the storage site for folates. A transport resistant line of the H35 cells has been developed and the properites of the cells will be compared with the normal H35 cells. Both hepatocytes and especially H35 cells catalyze a rapid conversion of methotrexate to polyglutamate derivatives. The properties of this reaction will be studied with cells in culture and cell free extracts to understand the enzymology and regulation of the reaction. We will also assess the contribution that the polyglutamates make to the chemotherapeutic action of methotrexate in H35 cells. Methotrexate bound to macromolecular carriers will be examined to determine their comparative effectiveness against normal and resistant H35 cells.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA025933-06
Application #
3167073
Study Section
Experimental Therapeutics Subcommittee 2 (ET)
Project Start
1979-07-01
Project End
1985-12-31
Budget Start
1985-01-01
Budget End
1985-12-31
Support Year
6
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
New York State Department of Health
Department
Type
DUNS #
002436061
City
Menands
State
NY
Country
United States
Zip Code
12204

  Publications

Yin, Dezhong; Galivan, John; Ao, Wei et al. (2003) Characterization of the human gamma-glutamyl hydrolase promoter and its gene expression in human tissues and cancer cell lines. Gene 312:281-8
Chave, Karen J; Ryan, Thomas J; Chmura, Stacey E et al. (2003) Identification of single nucleotide polymorphisms in the human gamma-glutamyl hydrolase gene and characterization of promoter polymorphisms. Gene 319:167-75
Cole, P D; Kamen, B A; Gorlick, R et al. (2001) Effects of overexpression of gamma-Glutamyl hydrolase on methotrexate metabolism and resistance. Cancer Res 61:4599-604
Sanchez-Alcazar, J A; Khodjakov, A; Schneider, E (2001) Anticancer drugs induce increased mitochondrial cytochrome c expression that precedes cell death. Cancer Res 61:1038-44
Sanchez-Alcazar, J A; Schneider, E; Martinez, M A et al. (2000) Tumor necrosis factor-alpha increases the steady-state reduction of cytochrome b of the mitochondrial respiratory chain in metabolically inhibited L929 cells. J Biol Chem 275:13353-61
Galivan, J; Ryan, T J; Chave, K et al. (2000) Glutamyl hydrolase. pharmacological role and enzymatic characterization. Pharmacol Ther 85:207-15
Chave, K J; Auger, I E; Galivan, J et al. (2000) Molecular modeling and site-directed mutagenesis define the catalytic motif in human gamma -glutamyl hydrolase. J Biol Chem 275:40365-70
Sanchez-Alcazar, J A; Ault, J G; Khodjakov, A et al. (2000) Increased mitochondrial cytochrome c levels and mitochondrial hyperpolarization precede camptothecin-induced apoptosis in Jurkat cells. Cell Death Differ 7:1090-100
Volk, E L; Rohde, K; Rhee, M et al. (2000) Methotrexate cross-resistance in a mitoxantrone-selected multidrug-resistant MCF7 breast cancer cell line is attributable to enhanced energy-dependent drug efflux. Cancer Res 60:3514-21
Galivan, J; Ryan, T; Rhee, M et al. (1999) Glutamyl hydrolase: properties and pharmacologic impact. Semin Oncol 26:33-7

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