Molecular analysis of the prostate-specific antigen upstream gene enhancer

Prostate. 2001 Jan 1;46(1):76-85. doi: 10.1002/1097-0045(200101)46:1<76::aid-pros1011>3.0.co;2-4.

Abstract

Background: Our objective was to identify factors other than androgen receptor that bind to and regulate the prostate-specific antigen (PSA) upstream gene enhancer (PSE).

Methods: DNAse I footprinting and electromobility shift assays (EMSA) were performed over the PSE using lysates from PSA-producing cell lines, LNCaP and LAPC4, and nonproducing PSA cell lines, PC-3 cells, U937 monocytes, and Namalwa B cells. Mutational analysis and transient transfection assays were used to determine the contributions made by different elements towards the regulation of the enhancer.

Results: Three distinct regions surrounding androgen response elements of the PSE were found to bind unknown ubiquitous and cell type-specific proteins. These regions, when mutated in a PSE reporter construct, were shown to be required for maximal activation in LNCaP cells.

Conclusions: These results correlate unknown sequence-specific DNA binding proteins with androgen-mediated regulation of a prostate-specific gene, thus providing further insight into the mechanism of PSA gene expression.

MeSH terms

  • Base Sequence
  • DNA Footprinting
  • DNA Primers / chemistry
  • Deoxyribonuclease I / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Enhancer Elements, Genetic / genetics*
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Male
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Plasmids / chemistry
  • Prostate-Specific Antigen / chemistry
  • Prostate-Specific Antigen / genetics*
  • Prostatic Neoplasms / chemistry
  • Prostatic Neoplasms / genetics*
  • Transfection
  • Tumor Cells, Cultured

Substances

  • DNA Primers
  • Deoxyribonuclease I
  • Prostate-Specific Antigen