Project description:UM1 and UM2 are paired cell lines generated from a oral cancer patient [Nakayama, et al Invas. Metast., 18 (1998), pp. 219–228]. UM1 is highly invasive and exhibits elevated metastatic potential as compared to UM2.
Project description:Ornithine decarboxylase antizyme 1 mediates DNA demethylation. Using a human oral cancer cell line, UM1, genes induiced by DNA demethylation were screened. To verify the methylation status of those induced genes, the gene expression profile was compared with that of 5-Aza-2’-deoxycytidine treated sample. Keywords: DNA demthylation, ornithine decarboxylase antizyme 1, 5-Aza-2’-deoxycytidine
Project description:High glucose has a significant effect on cancer progression. We aim to investigate the effect of glucose on oral cancer cells at gene expression level. We use microarray to identify differentially expressed gene in oral cancer cells (UM1) that cultured in different glucose concentration.
Project description:Ornithine decarboxylase antizyme 1 mediates DNA demethylation. Using a human oral cancer cell line, UM1, genes induiced by DNA demethylation were screened. To verify the methylation status of those induced genes, the gene expression profile was compared with that of 5-Aza-2â-deoxycytidine treated sample. Preparation of samples, hybridization process and data analysis were performed according to the manufacture's protocol (BD Biosciences). To verify the genes induced by ornithine decarboxylase antizyme via DNA demethylation, 5-Aza-2â-deoxycytidine treatment was carried out as a parallel experiment.
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
Project description:Kynureninase is a member of a large family of catalytically diverse but structurally homologous pyridoxal 5'-phosphate (PLP) dependent enzymes known as the aspartate aminotransferase superfamily or alpha-family. The Homo sapiens and other eukaryotic constitutive kynureninases preferentially catalyze the hydrolytic cleavage of 3-hydroxy-l-kynurenine to produce 3-hydroxyanthranilate and l-alanine, while l-kynurenine is the substrate of many prokaryotic inducible kynureninases. The human enzyme was cloned with an N-terminal hexahistidine tag, expressed, and purified from a bacterial expression system using Ni metal ion affinity chromatography. Kinetic characterization of the recombinant enzyme reveals classic Michaelis-Menten behavior, with a Km of 28.3 +/- 1.9 microM and a specific activity of 1.75 micromol min-1 mg-1 for 3-hydroxy-dl-kynurenine. Crystals of recombinant kynureninase that diffracted to 2.0 A were obtained, and the atomic structure of the PLP-bound holoenzyme was determined by molecular replacement using the Pseudomonas fluorescens kynureninase structure (PDB entry 1qz9) as the phasing model. A structural superposition with the P. fluorescens kynureninase revealed that these two structures resemble the "open" and "closed" conformations of aspartate aminotransferase. The comparison illustrates the dynamic nature of these proteins' small domains and reveals a role for Arg-434 similar to its role in other AAT alpha-family members. Docking of 3-hydroxy-l-kynurenine into the human kynureninase active site suggests that Asn-333 and His-102 are involved in substrate binding and molecular discrimination between inducible and constitutive kynureninase substrates.