Expression of the androgen-dependent MMTV-specific orf gene in Shionogi 115 mouse mammary tumor cells
| Autor: | Philippa D. Darbre, Pirkko Härkönen, Eeva M. Valve, Merja J. Tasanen, Johanna K. Ruohola, James F. Glover |
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| Rok vydání: | 2001 |
| Předmět: |
Male
medicine.medical_specialty DNA Complementary Neoplasms Hormone-Dependent Genes Viral viruses Endocrinology Diabetes and Metabolism Molecular Sequence Data Clinical Biochemistry Cell Gene Expression Mice Nude Biology Transfection Fibroblast growth factor Biochemistry Mice Open Reading Frames Endocrinology Complementary DNA Internal medicine Tumor Cells Cultured medicine Animals Amino Acid Sequence RNA Messenger Cloning Molecular Promoter Regions Genetic Molecular Biology Gene Mammary tumor Messenger RNA Base Sequence Sequence Homology Amino Acid Mammary Neoplasms Experimental 3T3 Cells Cell Biology Molecular biology medicine.anatomical_structure Mammary Tumor Virus Mouse Cell culture DNA Viral Androgens RNA Viral Molecular Medicine Female Cell Division |
| Zdroj: | The Journal of Steroid Biochemistry and Molecular Biology. 78:389-400 |
| ISSN: | 0960-0760 |
| Popis: | The Shionogi 115 (S115) mouse mammary tumor cells express the MMTV-specific 1.7 kb mRNA (orf) at a high level in the presence of androgens. In lymphoid cells the orf-gene encodes a superantigen which has an important role in establishing self-tolerance but in mammary and breast cancer cells the function of the orf gene is unclear. In the present work we studied the expression of the S115 mammary tumor cell orf sequence and its role in the androgen regulated growth of S115 cells. The cloning and sequencing of the cDNA specific for the 1.7 kb mRNA from the S115 mouse mammary tumor cells revealed a 990 bp DNA sequence with a 99.8% homology to the Mtv-17 proviral strain. There was a difference of only one amino acid (isoleu-tyr) in the coding region. A peptide was synthesized according to the hypervariable C-terminal part of the predicted protein and used to raise a rabbit antiserum. The anti-S115-orf antiserum immunoprecipitated an approximately 45 kDa protein from the metabolically labeled S115 cell lysates. In order to analyze the putative functions of the protein, the orf-sequence was linked to MoMLV-LTR and to the human s-actin promoter in the mammalian expression vectors pLTRpoly and pHsAPr-1-neo, respectively, and transfected into NIH3T3 and S115 cells. NIH3T3 transfectants expressing orf mRNA did not show a transformed phenotype in vitro. The S115 orf transfectants proliferated somewhat more slowly than the vector transfected control cells in cell culture, both in the presence or absence of androgen, but there was no obvious change in the phenotype of S115 cells or in expression of the fibroblast growth factor 8 (FGF-8). This factor is activated by Mtv-6 integration and mediates androgen effects in these cells. Unexpectedly, however, the formation of tumors by S115 orf cells in nude mice was considerably prolonged and tumor growth retarded when compared with vector transfected control or parent S115 cells. The results suggest that MMTV-orf can be functional in breast cancer cells but the mechanism of the growth repressive effect in mammary tumor remains to be analyzed. |
| Databáze: | OpenAIRE |
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