Androgen Receptor Gene Mutations in Human Prostate Cancer

• Published in: Proceedings of the National Academy of Sciences - PNAS 1992-07, Vol.89 (14), p.6319-6323
• Main Authors: Newmark, Jay R., Hardy, Dianne O., Tonb, Dalal C., Carter, Bob S., Epstein, Jonathan I., Isaacs, William B., Brown, Terry R., Barrack, evelyn R.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Androgens; Base Sequence; Binding Sites; Biochemistry; Biological and medical sciences; carcinoma; Cell lines; Codons; Exons; Genes; Genetic mutation; Genetics; Gynecology. Andrology. Obstetrics; Humans; In Vitro Techniques; Lymphocytes; Male; Male genital diseases; man; Medical sciences; Molecular Sequence Data; Mutation; Oligodeoxyribonucleotides - chemistry; Polymerase Chain Reaction; prostate; Prostate cancer; Prostatic Neoplasms - genetics; receptors; Receptors, Androgen - genetics; Tumor Cells, Cultured; Tumors
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.89.14.6319

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.