Mutational analysis of the androgen receptor using laser capture microdissection and direct sequencing.
Molecular analysis of prostate cancer specimens is complicated by tumor heterogeneity and admixture of malignant cells with benign epithelium, stroma and inflammatory cells. Laser Capture Microdissection allows procurement of selected groups of cells from specific areas of tissue sections. We have used Laser Capture Microdissection to obtain pure samples of malignant prostate epithelial cells from frozen radical prostatectomy specimens. Methods for DNA extraction, androgen receptor amplification and exon sequencing have been optimized. These methods should be adaptable for molecular analysis of DNA, RNA and protein from other complex tissues.
Greene
Sheila
S
Division of Urology, Department of Surgery, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Stockton
Patricia
P
Kozyreva
Olga G
OG
Gaston
Kris E
KE
Flake
Gordon P
GP
Mohler
James L
JL
D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D006801:Humans; D007834:Lasers; D008297:Male; D008846:Micromanipulation / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D011471:Prostatic Neoplasms / Q000235:genetics; D011944:Receptors, Androgen / Q000235:genetics*
D017931:DNA Primers; D011944:Receptors, Androgen