Expression data of mouse NIH3T3 and NIH3T3-k-ras transformed cell lines after Forskolin treatment
ABSTRACT: Forskolin treatment induces activation of protein kinase A. This activation protects transformed cells from death induced by glucose starvation. We used microarray to identify the mechanisms involved in such a pro-survival effect. NIH3T3 and NIH3T3-K-ras were plated at 1mM glucose and then daily treated or not with 10uM FSK until 72h of growth. The microarray analysis has been performed at 72h in order to identify specific mechanisms associate with transformed cells survival.
Project description:Expression profiling of normal NIH3T3 and transformed NIH3T3 K-ras cell lines grown for 72 hours in optimal glucose availability (25 mM glucose) or low glucose availability (1 mM). Low glucose induces apoptosis in transformed cells as compared to normal ones. We performed genome-wide analysis by using Affymetrix GeneChip oligonucleotide microarrays to identify those genes whose expression levels are modulated by glucose availability in NIH3T3 and NIH3T3 K-ras cell lines. The cells, to be used for the transcription analysis, were collected at 18h from the initial seeding, time corresponding to the change of medium (25 or 1 mM glucose, indicated as T0) and then at 24, 48 and 72h upon medium change.
Project description:The aim of this experiment was to get a comparison of the signatures between a non-transformed cell (NIH3T3 + vector) and a transformed cell (NIH3T3 + Fbxo7). NIH3T3 cells become transformed after the stable integration of the Fbxo7 gene. Fbxo7 potentiates cyclin D/cdk6 activity.
Project description:Effect of the overexpression of the oncogenic forms of the Vav2 and Vav proteins in the NIH3T3 cell line. oncovav- and oncovav2-transformed NIH3T3 cells are compared to the parental NIH3T3 controls under exponential growth conditions
Project description:High-temporal resolution profiling was performed on NIH3T3 fibroblasts to detect rhythmic transcripts Experiment Overall Design: Samples were collected every hour for 48 hours from forskolin-synchronized NIH3T3 cells. Samples were analyzed using Affymetrix arrays.
Project description:Effect of the overexpression of the oncogenic form of the Vav2 protein in the NIH3T3 cell line under serum deprivation conditions. oncovav2-transformed NIH3T3 cells grown in serum-deprived medium (Vav2SD) are compared to the parental NIH3T3 controls under the same growth conditions (ContSD). Vav2SD cells are also compared to the oncovav2-transformed NIH3T3 cells growing exponentially and the NIH3T3 growing exponentially.
Project description:Interferons have been ascribed to mediate antitumor effects. IRF-1 is a major target gene of interferons. It inhibits cell proliferation and oncogenic transformation. Here we show that 60% of all mRNAs deregulated by oncogenic transformation mediated by c-myc and H-ras are reverted to the expression levels of non-transformed cells by IRF-1. These include cell cycle regulating genes. Activation of IRF-1 decreases cyclin D1 expression and CDK4 kinase activity concomitant with dephosphorylation of pRb. These effects of IRF-1 are mediated by inhibition of the MEK-ERK pathway and a transcriptional repression of cyclin D1. IRF-1 mediated effects on cell cycle progression were found to be overridden by ectopic expression of cyclin D1. Ablation of cyclin D1 by RNA interference experiments prevents transformation and tumor growth in nude mice. The data demonstrate that cyclin D1 is a key target for IRF-1 mediated tumor suppressive effects. Experiment Overall Design: 3 samples were analysed, two replicates per sample. NIH3T3 cells were compared with myc/ras transformed NIH3T3 cells and with IRF1 expressing myc/ras transformed cells.
Project description:Transcriptional profiling of mouse NIH3T3 cells comparing control NIH3T3 cells transfected with a pEFm6-BRAF with cells transfected with pEFm6-BRAFV600E. Goal was to determine the effects of BRAFV600E gene transfection on global mouse NIH3T3 cells gene expression. Overall design: Two-condition experiment, Wild vs. BRAFV600E TransfectedNIH3T3 cells.
Project description:Polyamines are absolutely required for cell growth and proliferation. While polyamine depletion results in reversible cell cycle arrest, the actual mechanism of growth inhibition is still obscure. This work aimed at determining the cellular processes affected by reduction in the intracellular polyamine levels In order to reveal the general transcriptional responses to polyamine depletion in mammalian cells, NIH3T3 mouse fibroblasts were treated with 1mM L-Difluoromethylornithine (DFMO), G1 cellular fractions were collected by sorting at 0,12, 24, 48 and 96 hours upon addition of the reagent, total RNA was extracted, reverse-transcribed, fragmented, labeled and hybridized to Affymetrix MoGene 1.0 ST DNA array.
Project description:To evaluate whether FEAT has cellular functions relevant in oncogenesis, FEAT was overexpressed in NIH3T3 cells, which only weakly express FEAT protein, and the alterations in genome-wide transcriptional profiles were analyzed by microarrays. Overall design: The ORF of human FEAT cDNA was subcloned into the pENTR3C entry vector of the Gateway cloning system (Invitrogen). LR recombination with the pEF-DEST51 vector (Invitrogen) yielded the pEF-DEST51-FEAT plasmid that encodes FEAT driven by the elongation factor 1alpha promoter. NIH3T3 cells were transfected with pEF-DEST51-FEAT using HilyMax (Dojindo Laboratories). After 7 days of selection with 10 µg/ml blasticidin S (Kaken Pharmaceutical), colonies were picked and screened for clones in which FEAT protein was overexpressed. Three clones, FEAT-3, -15, and -20 were used in further studies. The ccdB gene in the pEF-DEST51 vector was deleted to construct the pEF-DEST51-∆ccdB plasmid, which was stably transfected into NIH3T3 cells to obtain control cell lines, named ∆ccdB-1, -2, and -3.
Project description:Identification of cyclical expressed coding and non-coding genes during the circadian rhythm in NIH3T3 cells. NIH3T3 cells were synchronized for their circadian rhythm and RNA sequencing were performed at several time points along the rhythm. This data was used to identify cyclical expressed genes as well as long intergenic non-coding RNAs. NIH3T3 cells were synchronized with 100 nM Dexamethasone for 2 hours, then medium was changed to normal culture medium (0h). Every 4 hours cells were harvested, RNA isolated and RNAseq performed.