Expression and function of normal and LNCaP androgen receptors in androgen-insensitive human prostatic cancer cells. Altered hormone and antihormone specificity in gene transactivation.
The gene for the androgen receptor (AR) in the androgen-sensitive human prostate cancer cell line LNCaP has a single-base mutation that produces a threonine to alanine change in the androgen-binding domain. Androgen-insensitive prostatic cancer (PC-3) cells were cotransfected with an expression vector encoding normal, LNCaP, or chimeric normal/LNCaP AR and a vector carrying a chloramphenicol acetyltransferase (CAT) reporter gene linked to the mouse mammary tumor virus promoter. CAT activity was specifically induced by androgens in PC-3 cells expressing normal AR. In PC-3 cells expressing LNCaP AR, however, CAT activity was also induced by progestins and the antiandrogen hydroxyflutamide, which had little activity in cells expressing normal AR. Steroid-binding competition assays using in vitro synthesized ARs showed that LNCaP AR had a higher affinity than normal AR for progestins, 17 beta-estradiol, and hydroxyflutamide. The antiprogestin and antiglucocorticoid RU 38486 induced CAT activity in PC-3 cells expressing normal AR but not LNCaP AR. These studies indicate that AR mutations may be very important in determining the appropriate method of treatment with steroid hormones or their antagonists.
Kokontis
J
J
Ben May Institute, University of Chicago, IL 60637.
Ito
K
K
Hiipakka
R A
RA
Liao
S
S
D000726:Androgen Antagonists / Q000494:pharmacology; D000728:Androgens / Q000378:metabolism / Q000502:physiology*; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D008297:Male; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics; D015735:Mifepristone / Q000494:pharmacology; D011471:Prostatic Neoplasms / Q000737:chemistry* / Q000473:pathology / Q000503:physiopathology; D014176:Protein Biosynthesis / Q000235:genetics; D011944:Receptors, Androgen / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D012016:Reference Values; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection / Q000235:genetics; D014407:Tumor Cells, Cultured
D000726:Androgen Antagonists; D000728:Androgens; D011944:Receptors, Androgen; D015735:Mifepristone; D015500:Chloramphenicol O-Acetyltransferase