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Alterations in cell growth and signaling in ErbB3 binding protein-1 (Ebp1) deficient mice.

Zhang Y, Lu Y, Zhou H, Lee M, Liu Z, Hassel BA, Hamburger AW - BMC Cell Biol. (2008)

Bottom Line: The proliferation of fibroblasts derived from Day 12.5 knock out embryos was also decreased as compared to that of wild type cells.Microarray expression analysis revealed changes in genes important in cell growth including members of the MAPK signal transduction pathway.These results indicate that Ebp1 can affect growth in an animal model, but that the expression of proliferation related genes is cell and context specific.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, University of Maryland School of Medicine, Baltimore, MD, USA. yzhan001@umaryland.edu

ABSTRACT

Background: The ErbB3 binding protein-1 (Ebp1) belongs to a family of DNA/RNA binding proteins implicated in cell growth, apoptosis and differentiation. However, the physiological role of Ebp1 in the whole organism is not known. Therefore, we generated Ebp1-deficient mice carrying a gene trap insertion in intron 2 of the Ebp1 (pa2g4) gene.

Results: Ebp1-/- mice were on average 30% smaller than wild type and heterozygous sex matched littermates. Growth retardation was apparent from Day 10 until Day 30. IGF-1 production and IGBP-3 and 4 protein levels were reduced in both embryo fibroblasts and adult knock-out mice. The proliferation of fibroblasts derived from Day 12.5 knock out embryos was also decreased as compared to that of wild type cells. Microarray expression analysis revealed changes in genes important in cell growth including members of the MAPK signal transduction pathway. In addition, the expression or activation of proliferation related genes such as AKT and the androgen receptor, previously demonstrated to be affected by Ebp1 expression in vitro, was altered in adult tissues.

Conclusion: These results indicate that Ebp1 can affect growth in an animal model, but that the expression of proliferation related genes is cell and context specific. The Ebp1-/- mouse line represents a new in vivo model to investigate Ebp1 function in the whole organism.

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Changes in expression of Ebp1 target genes in adult tissue. A. Number 4 mammary glands were isolated from 10 week old wild type (WT) or Ebp1 knock out (KO) mice. Immunohistochemical analysis was performed for phospho AKT (p-AKT) or total AKT (T-AKT) as indicated. B. Expression of AR and AR regulated genes in prostates. The expression of AGR2 in the anterior prostates of age matched wild type and Ebp1 knock out mice were measured by real-time PCR (left panel) or Western blot (middle panel) analysis. AR levels were measured in lysates of anterior prostates by Western blot analysis (Right panel).
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Figure 7: Changes in expression of Ebp1 target genes in adult tissue. A. Number 4 mammary glands were isolated from 10 week old wild type (WT) or Ebp1 knock out (KO) mice. Immunohistochemical analysis was performed for phospho AKT (p-AKT) or total AKT (T-AKT) as indicated. B. Expression of AR and AR regulated genes in prostates. The expression of AGR2 in the anterior prostates of age matched wild type and Ebp1 knock out mice were measured by real-time PCR (left panel) or Western blot (middle panel) analysis. AR levels were measured in lysates of anterior prostates by Western blot analysis (Right panel).

Mentions: Previous in vitro work with Ebp1 has used breast and prostate cancer cell lines derived from adults. In these tissues, the expression of Cyclin D1[24], AGR2 [27] and androgen receptor [14] have been shown to be regulated by Ebp1. We therefore examined expression of these genes in adult mammary and prostate epithelium. We found that in mammary epithelial cells, Cyclin D1 protein levels were not changed in knock out mice as compared to wild type (data not shown). However, the phosphorylation of AKT was greatly increased in Ebp1-/- mammary tissue (Fig. 7A). In prostate cancer cell lines, the expression of the androgen receptor and the metastasis associated protein AGR2 has been demonstrated to be decreased by ectopic expression of Ebp1. We found that AGR2 mRNA (Fig. 7B, right panel) and protein (Fig. 7B, middle panel) expression and AR protein expression (Fig. 7B, right panel) were increased in prostates of Ebp1 knock out mice as compared to age matched controls.


Alterations in cell growth and signaling in ErbB3 binding protein-1 (Ebp1) deficient mice.

Zhang Y, Lu Y, Zhou H, Lee M, Liu Z, Hassel BA, Hamburger AW - BMC Cell Biol. (2008)

Changes in expression of Ebp1 target genes in adult tissue. A. Number 4 mammary glands were isolated from 10 week old wild type (WT) or Ebp1 knock out (KO) mice. Immunohistochemical analysis was performed for phospho AKT (p-AKT) or total AKT (T-AKT) as indicated. B. Expression of AR and AR regulated genes in prostates. The expression of AGR2 in the anterior prostates of age matched wild type and Ebp1 knock out mice were measured by real-time PCR (left panel) or Western blot (middle panel) analysis. AR levels were measured in lysates of anterior prostates by Western blot analysis (Right panel).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2648959&req=5

Figure 7: Changes in expression of Ebp1 target genes in adult tissue. A. Number 4 mammary glands were isolated from 10 week old wild type (WT) or Ebp1 knock out (KO) mice. Immunohistochemical analysis was performed for phospho AKT (p-AKT) or total AKT (T-AKT) as indicated. B. Expression of AR and AR regulated genes in prostates. The expression of AGR2 in the anterior prostates of age matched wild type and Ebp1 knock out mice were measured by real-time PCR (left panel) or Western blot (middle panel) analysis. AR levels were measured in lysates of anterior prostates by Western blot analysis (Right panel).
Mentions: Previous in vitro work with Ebp1 has used breast and prostate cancer cell lines derived from adults. In these tissues, the expression of Cyclin D1[24], AGR2 [27] and androgen receptor [14] have been shown to be regulated by Ebp1. We therefore examined expression of these genes in adult mammary and prostate epithelium. We found that in mammary epithelial cells, Cyclin D1 protein levels were not changed in knock out mice as compared to wild type (data not shown). However, the phosphorylation of AKT was greatly increased in Ebp1-/- mammary tissue (Fig. 7A). In prostate cancer cell lines, the expression of the androgen receptor and the metastasis associated protein AGR2 has been demonstrated to be decreased by ectopic expression of Ebp1. We found that AGR2 mRNA (Fig. 7B, right panel) and protein (Fig. 7B, middle panel) expression and AR protein expression (Fig. 7B, right panel) were increased in prostates of Ebp1 knock out mice as compared to age matched controls.

Bottom Line: The proliferation of fibroblasts derived from Day 12.5 knock out embryos was also decreased as compared to that of wild type cells.Microarray expression analysis revealed changes in genes important in cell growth including members of the MAPK signal transduction pathway.These results indicate that Ebp1 can affect growth in an animal model, but that the expression of proliferation related genes is cell and context specific.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, University of Maryland School of Medicine, Baltimore, MD, USA. yzhan001@umaryland.edu

ABSTRACT

Background: The ErbB3 binding protein-1 (Ebp1) belongs to a family of DNA/RNA binding proteins implicated in cell growth, apoptosis and differentiation. However, the physiological role of Ebp1 in the whole organism is not known. Therefore, we generated Ebp1-deficient mice carrying a gene trap insertion in intron 2 of the Ebp1 (pa2g4) gene.

Results: Ebp1-/- mice were on average 30% smaller than wild type and heterozygous sex matched littermates. Growth retardation was apparent from Day 10 until Day 30. IGF-1 production and IGBP-3 and 4 protein levels were reduced in both embryo fibroblasts and adult knock-out mice. The proliferation of fibroblasts derived from Day 12.5 knock out embryos was also decreased as compared to that of wild type cells. Microarray expression analysis revealed changes in genes important in cell growth including members of the MAPK signal transduction pathway. In addition, the expression or activation of proliferation related genes such as AKT and the androgen receptor, previously demonstrated to be affected by Ebp1 expression in vitro, was altered in adult tissues.

Conclusion: These results indicate that Ebp1 can affect growth in an animal model, but that the expression of proliferation related genes is cell and context specific. The Ebp1-/- mouse line represents a new in vivo model to investigate Ebp1 function in the whole organism.

Show MeSH
Related in: MedlinePlus