Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse mock treated L929 cells with L929 cells infected with wildtype versus nt3A mutant VRP at 5hpi

ABSTRACT: This series includes arrays from two separate experiments where the gene expression profile of mock treated L929 cells was compared to that of L929 cells that had been infected with wildtype versus nt3A mutant Venezuelan equine encephalitis virus replicon particles (VRP). Total RNA was isolated from all samples at 5 hour post-infection. Experiment Overall Design: L929 cells were mock treated or infected at an MOI of 5 with either wildtype or nt3A mutant null-VRP, and total RNA was isolated at 5 hpi using the UltraSpec reagent protocol as described above. The UNC-CH Functional Genomics Core Facility provided Affymetrix sample preparation and hybridization services, hybridizing the samples to Affymetrix Mouse Expression 430A (MOE430A) GeneChip arrays, which include analysis of ~22,600 genes. Affymetrix GeneChip Microarray Suite 5.0 software was used for washing, scanning, and basic analysis, with sample quality assessed by examination of 3â to 5â intensity ratios of certain genes. Expression analysis files created by Microarray Suite were imported into GeneSpring 6.2 (Silicon Genetics) for further gene expression analysis and filtering. Two independent array studies were completed, and gene lists were compiled representing only those genes that were up- or downregulated by greater than 2-fold in replicate analyses.

ORGANISM(S): Mus musculus

SUBMITTER: Jennifer Konopka

PROVIDER: E-GEOD-7647 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Sialic acids on vertebrate cell surfaces mediate many biological roles. Altered expression of certain sialic acid types or their linkages can have prognostic significance in human cancer. A classic but unexplained example is enhanced Î±2-6-sialylation on N-glycans, resulting from over-expression of the Golgi enzyme Î²-galactoside:Î±2-6-sialyltransferase (ST6Gal-I). Previous data supporting a role for the resulting SiaÎ±2-3GalÎ²1-4GlcNAc (Sia6LacNAc) structure in tumor biology were based on in vitro studies in transfected carcinoma cells, in which increased Sia6LacNAc on Î²1-integrins enhanced their binding to ligands, and stimulated cell motility. Here we examine for the first time the in vivo role of the ST6Gal-I enzyme in the growth and differentiation of spontaneous mammary cancers in mice transgenic for an MMTV-promoter-driven polyoma-middle-T antigen, a tumor in which beta1-integrin function is important for tumorigenesis, and in maintaining the proliferative state of tumor cells. Tumors induced in St6gal1 null animals were more differentiated in comparison to those in the wild-type background, both by histological analysis and by protein expression profiles. Furthermore, we show the St6gal1 null tumors have selectively altered expression of genes associated with focal adhesion signaling, and have decreased phosphorylation of FAK, a downstream target of Î²1-integrins. This first in vivo evidence for a role of ST6Gal-I in tumor progression was confirmed using a novel approach, which conditionally restored St6gal1 in cell lines derived from the null tumors. These findings indicate a role for ST6Gal-I as a mediator of tumor progression, with its expression causing a less differentiated phenotype, via enhanced Î²1-integrin function. Experiment Overall Design: Messenger RNA from tumors arising in wild-type or St6gal1 null littermates was used for the gene expression analysis on the Mouse Genome 430 2.0 Array (Affymetrix). Six PyV-mT St6gal1 null and wild-type tumor pairs were used, in four independent pools of three tumors each. Each sample is a pool of three different tumors from three different animals. For the first set of microarray analysis samples GSM239969 and GSM239970 were being compared, and in the second set, samples GSM239971 and GSM239972 were compared.

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Transcription profiling of mouse mock treated L929 cells with L929 cells infected with wildtype versus nt3A mutant VRP at 5hpi

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