title: Androgen receptor gene mutations in human prostate cancer.
abstract: We screened human prostate cancer tissues for the presence of somatic mutations in the hormone binding domain of the androgen receptor (AR) gene. Exons E-H were amplified from genomic DNA using the polymerase chain reaction and analyzed by denaturing gradient gel electrophoresis (DGGE), which separates DNA fragments that differ by only a single base. We detected a mutation in exon E of the hormone binding domain in 1 of 26 specimens of untreated organ-confined stage B prostate cancer. The mutation was not detectable in peripheral blood lymphocyte DNA. Lymphocyte DNA (wild-type AR) migrated in DGGE as a single band. The tumor DNA migrated in DGGE as four bands, consistent with the presence of cells with mutant AR plus cells with wild-type AR and indicating that the tumor contained a somatic mutation. To our knowledge, a somatic AR gene mutation has not been reported previously. Sequencing revealed a G----A substitution in codon 730, changing valine to methionine. Codon 730 is in a region highly conserved among all steroid receptors. The abundance of the mutated fragment (about 50% of the DNA in the specimen) indicates its presence in cells with a growth advantage. A somatic mutation could be detected by DGGE if it represented at least 10% of the sample. Failure to detect mutations in other specimens analyzed may be due to this limit of sensitivity, the presence of mutations in other parts of the AR, or a low frequency of mutations in early stage disease.
authors.affiliation: Department of Urology, Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: J R
authors.initials: JR
authors.lastname: Newmark
authors.forename: D O
authors.initials: DO
authors.lastname: Hardy
authors.forename: D C
authors.initials: DC
authors.lastname: Tonb
authors.forename: B S
authors.initials: BS
authors.lastname: Carter
authors.forename: J I
authors.initials: JI
authors.lastname: Epstein
authors.forename: W B
authors.initials: WB
authors.lastname: Isaacs
authors.forename: T R
authors.initials: TR
authors.lastname: Brown
authors.forename: E R
authors.initials: ER
authors.lastname: Barrack
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011471:Prostatic Neoplasms / Q000235:genetics*; D011944:Receptors, Androgen / Q000235:genetics*; D014407:Tumor Cells, Cultured
chemical_list: D009838:Oligodeoxyribonucleotides; D011944:Receptors, Androgen