Differential gene expression pattern of mouse livers transfected with dual oncogenes (day-7 post-injection data set)
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ABSTRACT: To understand global gene expression patterns of mouse livers upon dual-oncogene transfection, RNA sequencing and analysis were conducted on liver samples from Shp2F/F or Shp2∆H (Shp2F/F:Alb-cre) mice hydrodynamically injected with the combination of Met/β-Catenin or Met/PIK3CA .
Project description:To understand global gene expression patterns of mouse livers upon dual-oncogene transfection, RNA sequencing and analysis were conducted on liver samples from Shp2F/F or Shp2∆H (Shp2F/F:Alb-cre) mice hydrodynamically injected with the combination of Met/β-Catenin or Met/PIK3CA .
Project description:Microarray analysis of WT (Pten2fl/fl:Shp2fl/fl:Alb-Cre-), SKO (Shp2hep-/-, or Shp2fl/fl:Alb-Cre+), PKO (Ptenhep-/-, or Pten2fl/fl:Alb-Cre+) and DKO (Ptenfl/fl:Shp2fl/fl:Alb-Cre+) liver samples to gain global molecular insights how shp2 and pten is involved in liver tumorigenesis.
Project description:To identify HGF/Met regulated genes, we performed expression microarray analysis after inducible activation of Met receptor in primary cultures of hepatocytes established from wild-type control (Alb-Cre +/-) and Met conditional knockout mice (Alb-Cre +/-; Met Fl/Fl). Keywords: time series design
Project description:To identify HGF/Met regulated genes, we performed expression microarray analysis after inducible activation of Met receptor in primary cultures of hepatocytes established from wild-type control (Alb-Cre +/-) and Met conditional knockout mice (Alb-Cre +/-; Met Fl/Fl). Total RNA was isolated from untreated hepatocyte cultures as well as from cultures treated with 50 ng/ml of HGF for 0.5, 2, 12 or 24 hours. RNA collected from these experiments was converted to fluorescently labeled cDNA and used for hybridizations of oligonucleotide microarrays. All experiments were repeated in triplicates. Total RNA from pooled wild-type mouse hepatocytes was used as universal reference and all hybridizations were repeated following a reverse flourescing.
Project description:The introduction of the tumorigenic v-Ha-ras oncogene-transformed rat liver epithelial cells (WBras), which is deficient in gap junctional intercellular communication (GJIC), into F344 rats, induces significant formation of hepatocellular tumors. GJIC plays a major role in maintaining tissue homeostasis. Using this in vivo tumor model system, we used 2-dimensional electrophoresis with isoelectric focusing in the first dimension and SDS-PAGE in the second dimension to globally identify proteins that are uniquely expressed in the livers of WBras-treated rats as compared to the sham control. Immunoblotting was used to identify Ras and Connexin43, which were the positive and negative marker proteins, respectively, of the introduced WBras cells. As predicted, immunoblotting indicated that the whole liver of tumor-bearing animals exhibited a decreased level of Connexin43 and an increased level of Ras. Connexin43 and GJIC were expressed and functional in normal liver, but not in the tumor. In addition to these 2 markers, an additional 4 proteins exhibited decreased levels and 2 proteins exhibited increased levels in the livers of tumor-bearing animals. N-Terminal sequencing analysis was used to identify these proteins, which were glucose-regulated protein 78, 2 isoforms of heat shock protein 60, and the beta-chain of ATP synthase for the down regulated proteins, and beta-Actin with a 46 amino acid deletion from its N-terminus and Vimentin with a 71 amino acid deletion from its N-terminus for the up regulated proteins. These data offer potentially new markers of liver tumorigenicity, particularly, Vimentin. (
Project description:To elucidate the functional roles of sMafs in the adult liver, we conditionally targeted the sMaf genes using a transgenic complementation rescue approach. MafF-/-::MafG-/-::MafK-/- (F0G0K0) mice are embryonic lethal but can be rescued by complementation of transgenic MafG expression under the regulation of the MafG regulatory domain (MGRD). Therefore, we rescued F0G0K0 mice using a MGRD transgenic mouse line with a MafG gene flanked with loxP (fMafG) sequences so that the MafG gene could be deleted by Cre-mediated recombination. The Albumin(Alb)-Cre transgenic mice were used to delete fMafG gene specifically in the liver. The genotype used are MafF-/-::MafG-/-::MafK-/-::MGRD-fMafG::Alb-Cre (liver-specific sMaf CKO) and MafF-/-::MafG+/-::MafK-/-::MGRD-fMafG::Alb-Cre (control).
Project description:In order to further discover the resistance mechanism of Rack1F/F;Alb-cre mice to LPS/ GalN-induced fulfulant hepatitis, we used genome-wide microarray expression profiling as a discovery platform to identify potential genes associated with resistance to LPS/ GalN-induced in Rack1F/F;Alb-cre mice. Fulminant hepatitis was induced by LPS/GalN in Rack1F/F;Alb-cre mice and Rack1F/F mice for 0, 1,3 and 6 hours, respectively.