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Estrogen receptor α (ESR1) over-expression mediated apoptosis in Hep3B cells by binding with SP1 proteins
- Source :
- Journal of molecular endocrinology. 51(1)
- Publication Year :
- 2013
-
Abstract
- Previous studies have reported that estrogen receptors (ERs) are expressed in normal human liver, chronic hepatitis, and benign hepatic tumor tissues. However, decreased expression of ERs can be observed in hepatocellular carcinoma (HCC) and the role of ERs in HCC is not fully understood. Thus, the present study aimed to investigate the molecular mechanism induced by the overexpression of ERα (ERα (ESR1)) in Hep3B cells. We first detected the induction of apoptosis in ER-negative Hep3B cells using DNA fragmentation assay and flow cytometry. We found that ERα and ERα plus 17β-estradiol treatment increased apoptosis in Hep3B cells. Additionally, western blotting showed increased expression of active caspase 3 and tumor necrosis factor α (TNFα (TNF)) inERα-transfected cells. To further understand the importance of SP1-binding sites in theTNFαpromoter, ERα-negative Hep3B cells were co-transfected withERαand a wild-type TNFα plasmid orTNFαwith deleted SP1 regions. Deletion of both distant and primal SP1 sites abolished the activity of ERα, and similar results were observed by blocking the expression of SP1 protein using mithramycin (MA). This result indicates that SP1 protein is essential for ERα-activatedTNFαpromoter activity. Co-immunoprecipitation assay further confirmed the binding interaction between ERα and SP1 in a ligand-dependent manner. In general, we demonstrate that the overexpression of ERα mediates apoptosis in ERα-negative Hep3B cells by the binding of ERα to SP1 protein. Additionally, this ERα–SP1 complex binds to the proximal and distal sites of theTNFαgene promoter and further induces the expression of active caspase 3 in a ligand-dependent manner.
- Subjects :
- Inhibitor of apoptosis domain
Sp1 transcription factor
Carcinoma, Hepatocellular
Transcription, Genetic
Chemistry
Sp1 Transcription Factor
Tumor Necrosis Factor-alpha
Liver Neoplasms
Estrogen Receptor alpha
Estrogen receptor
Gene Expression
Caspase 3
Apoptosis
Molecular biology
Gene Expression Regulation, Neoplastic
Endocrinology
Cell Line, Tumor
DNA fragmentation
Humans
Tumor necrosis factor alpha
Promoter Regions, Genetic
Molecular Biology
Estrogen receptor alpha
Estrogen receptor beta
Protein Binding
Subjects
Details
- ISSN :
- 14796813
- Volume :
- 51
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Journal of molecular endocrinology
- Accession number :
- edsair.doi.dedup.....1b6ea9dda8c40baaa43eda33bd721ee1