Project description:The human glucocorticoid receptor (GRα) is overexpressed at the molecular and protein level in malignant human adrenocortical cancers. A stable cell line model of GRα overexpression was established using the H295R human adrenocortical cancer cell line. The following results were obtained from gene expression profiling of H295R_GRα and H295R_Control (empty vector) cells following treatment with either a GRα agonist (dexamethasone), GRα antagonist (RU486) or vehicle (ethanol) control. H295R_GRα and H295R_Control (empty vector) cells were treated in triplicate for 6 hours with dexamethasone 100 nM, RU486 100 nM or vehicle (ethanol) control.
Project description:The human glucocorticoid receptor (GRα) is overexpressed at the molecular and protein level in malignant human adrenocortical cancers. A stable cell line model of GRα overexpression was established using the H295R human adrenocortical cancer cell line. The following results were obtained from gene expression profiling of H295R_GRα and H295R_Control (empty vector) cells following treatment with either a GRα agonist (dexamethasone), GRα antagonist (RU486) or vehicle (ethanol) control.
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:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.
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.
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. One-condition experment, gene expression of 3A6