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Involvement of 5-lipoxygenase metabolites of arachidonic acid in cyclic AMP-stimulated steroidogenesis and steroidogenic acute regulatory protein gene expression
Involvement of 5-lipoxygenase metabolites of arachidonic acid in cyclic AMP-stimulated steroidogenesis and steroidogenic acute regulatory protein gene expression
- Source :
- The Journal of Steroid Biochemistry and Molecular Biology. 85:159-166
- Publication Year :
- 2003
- Publisher :
- Elsevier BV, 2003.
-
Abstract
- To understand the mechanism for the role of arachidonic acid (AA) in steroidogenic acute regulatory (StAR) gene transcription, sections of the -1/-966 StAR promoter were deleted to produce constructs of -1/-426,-1/-211, -1/-151, and -1/-110 and inserted into the PGL3 vector to drive luciferase expression. Results indicated that -1/-151 StAR promoter contains the elements that are most responsive to AA. Electrophoretic mobility shift assays using nuclear extracts from AA-treated MA-10 Leydig tumor cells showed that AA enhanced specific binding of the nuclear extract to a 30 bp (-67/-96) sequence of the StAR promoter. Also, HPLC was used to identify AA metabolites involved in StAR gene transcription. It was found that 1 mM N6,2-O-dibutyryladenosine 3:5-cyclic monophosphate (dbcAMP) significantly increased the 5-lipoxygenase metabolites, 5-hydroperoxyeicosatetraenoic acid (5-HPETE) and 5-hydroxyeicosatetraenoic acid (5-HETE). Moreover, in the presence of 0.2 mM dbcAMP addition of 20 μM 5-HPETE or 5-HETE significantly enhanced StAR protein expression and progesterone production (P < 0.05). Similar results were obtained for StAR gene transcription with StAR mRNA levels and StAR promoter activities being significantly increased ( P < 0.05) when 5-HPETE was added to MA-10 cell cultures. In summary, the present studies demonstrated that cyclic AMP (cAMP) stimulated the production of the AA metabolites, 5-HPETE and 5-HETE, and showed that these metabolites enhanced StAR gene expression and steroid hormone production. The results further suggested that the AA-responsive element resides in the -67/-96 region of the StAR promoter.
- Subjects :
- endocrine system
Endocrinology, Diabetes and Metabolism
medicine.medical_treatment
Clinical Biochemistry
Arachidonic Acids
Biochemistry
Mice
chemistry.chemical_compound
Endocrinology
Genes, Reporter
Hydroxyeicosatetraenoic Acids
Gene expression
Cyclic AMP
Tumor Cells, Cultured
medicine
Animals
Luciferase
Luciferases
Promoter Regions, Genetic
Molecular Biology
DNA Primers
Sequence Deletion
Ovarian Neoplasms
Arachidonate 5-Lipoxygenase
Base Sequence
biology
Steroidogenic acute regulatory protein
Membrane Proteins
Cell Biology
Phosphoproteins
Molecular biology
Recombinant Proteins
Kinetics
Mutagenesis, Insertional
Steroid hormone
Bucladesine
Gene Expression Regulation
chemistry
Mrna level
Cell culture
Arachidonate 5-lipoxygenase
biology.protein
Molecular Medicine
Female
Arachidonic acid
Leydig Cell Tumor
Subjects
Details
- ISSN :
- 09600760
- Volume :
- 85
- Database :
- OpenAIRE
- Journal :
- The Journal of Steroid Biochemistry and Molecular Biology
- Accession number :
- edsair.doi.dedup.....e3315dc700aee71e38df336b7b91e567
- Full Text :
- https://doi.org/10.1016/s0960-0760(03)00189-4