Identification of estrogen regulated genes in Fe33 rat hepatoma cells by differential display polymerase chain reaction and their hormonal regulation in rat liver and uterus

Autor: A. Walter, P. Diel, K. H. Fritzemeier, Ch. Hegele-Hartung, Rudolf Knauthe
Rok vydání: 1995
Předmět:
medicine.medical_specialty
medicine.drug_class
Ovariectomy
Endocrinology
Diabetes and Metabolism
Molecular Sequence Data
Clinical Biochemistry
Estrogen receptor
Biology
Ethinyl Estradiol
Polymerase Chain Reaction
Biochemistry
Glucagon
Dexamethasone
Liver Neoplasms
Experimental
S100 Calcium Binding Protein G
Endocrinology
Internal medicine
Gene expression
Tumor Cells
Cultured
medicine
Animals
RNA
Messenger
Rats
Wistar
Molecular Biology
Progesterone
Messenger RNA
Differential display
Base Sequence
Estradiol
Uterus
Cell Biology
Blotting
Northern
Rats
Gene Expression Regulation
Neoplastic
Insulin-Like Growth Factor Binding Protein 1
Tamoxifen
Liver
Receptors
Estrogen
Estrogen
Ovariectomized rat
Molecular Medicine
Female
hormones
hormone substitutes
and hormone antagonists
Acute-Phase Proteins
Hormone
Zdroj: The Journal of Steroid Biochemistry and Molecular Biology. 55:363-373
ISSN: 0960-0760
DOI: 10.1016/0960-0760(95)00186-7
Popis: We applied the differential display RT-PCR (ddRT-PCR) technology to identify estrogen-regulated hepatic genes in the estrogen receptor expressing rat hepatoma cell line Fe33. Three genes of known sequences were detected by the ddRT-PCR approach: IGF binding protein-1 (IGFBP-1), vitamin D-dependent calcium-binding protein (CaBP9k) and major acute phase protein (MAP). Effects of ethinyl estradiol on the mRNA levels of these genes were confirmed by “Northern-blot” analysis. If given in combination with dexamethasone and glucagon, ethinyl estradiol caused 40-, 15- and 11-fold increases in the mRNA steady state level of IGFBP-1, CaBP9k and MAP, respectively, in Fe33 cells 24 h after addition of hormone. Besides ethinyl estradiol, the partial estrogen agonist OH-tamoxifen caused dose dependent effects on expression of MAP and IGFBP-1. Estrogen regulation of the respective genes and the modulatory effects of progesterone (10 mg/animal/day) were studied in ovariectomized rats treated subcutaneously for 14 days with 1 μg/animal/day estradiol. “Northern-blot” analysis of liver RNA revealed a 6-fold stimulation of IGFBP-1 mRNA levels in estradiol-treated compared to vehicle-treated rats and a weak but detectable increase of MAP mRNA steady state level (1.6-fold) upon estradiol administration. No effect of estradiol treatment could be monitored for CaBP9k in rat liver. Modulatory effects of progesterone on estradiol-stimulated expression in the liver could be monitored for IGFBP-1 only. In an extension of our investigation on the expression of the three genes in rat liver, we determined their expression and hormonal regulation in the uterus of the same animals. In the uterus, estradiol caused an increase in CaBP9k mRNA. In contrast, IGFBP-1 mRNA levels increased dramatically upon progesterone administration, whereas no effect of estradiol treatment could be detected. MAP mRNA levels increased only after coadministration of estradiol and progesterone. In conclusion, the ddRT-PCR proved to be a powerful method to identify estrogen-regulated genes. The study on the hormonal regulation of three genes stimulated by estrogen in Fe33 cells revealed similarities and differences in their regulationin vivo andin vitro.
Databáze: OpenAIRE
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