Induction of bicalutamide sensitivity in prostate cancer cells by an epigenetic Puralpha-mediated decrease in androgen receptor levels.
BACKGROUND
Increased androgen receptor (AR) levels support resistance to apoptosis and hormone therapy in advanced prostate cancer (PC). We recently linked the overexpression of AR in androgen-independent LNCaP cells (AI-cells) and tissues from castration-resistant patients to decreased nuclear levels of Pur-alpha (Puralpha) and loss from a protein complex bound to repressor sequences (ARS) in the 5'-UTR of AR. Strategies to regain control of increased AR transcription may overcome resistance of AI-cells and improve treatment outcomes.


METHODS
MTT, real-time PCR, Western blot, ChIP, flow cytometry, and caspase 3/7 activation measured the effect on growth and targets of LBH589/bicalutamide treatment of AI-cells and androgen-dependent LNCaP cells (AD).


RESULTS
Within 16 hr of treatment of AI-cells with low concentrations of the histone deacetylase inhibitor LBH589, a shift of cytoplasmic Puralpha restored the nuclear levels and the binding of Puralpha to the ARS. This was followed by a decline in AR-mRNA and protein reaching levels of parental AD-cells. The fraction of AI-cells in G1 increased and the cells in S phase decreased similar to AD-cells, and there was a modest caspase activation. Most notably, treatment of bicalutamide-resistant AI-cells with 10 nM LBH589 combined with 12.5 microM bicalutamide synergistically inhibited cell growth and induced a fivefold higher level of caspase 3/7 activation than observed in AD-cells.


CONCLUSIONS
Low-dose LBH589 restores Puralpha binding to ARS and down-regulates AR transcription. Biologically, LBH589 reverses the resistance of AI-cells to bicalutamide and to apoptosis. The combination may restore the hormonal response of castration-resistant PC patients.
New York University Cancer Institute and Department of Medicine, New York University School of Medicine, New York, New York 10016, USA.
Xiaomei
X
Liu
Alejandro
A
Gomez-Pinillos
Xiaojun
X
Liu
Edward M
EM
Johnson
Anna C
AC
Ferrari
D020121:5' Untranslated Regions; D000726:Androgen Antagonists / Q000494:pharmacology*; D000813:Anilides / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000378:metabolism*; D044127:Epigenesis, Genetic / Q000187:drug effects; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D007211:Indoles; D008297:Male; D009570:Nitriles / Q000494:pharmacology*; D009919:Orchiectomy; D000077767:Panobinostat; D011471:Prostatic Neoplasms; D011944:Receptors, Androgen / Q000096:biosynthesis / Q000235:genetics*; D014105:Tosyl Compounds / Q000494:pharmacology*; D014157:Transcription Factors / Q000378:metabolism*
D020121:5' Untranslated Regions; D000726:Androgen Antagonists; D000813:Anilides; D004268:DNA-Binding Proteins; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D007211:Indoles; D009570:Nitriles; C501130:PURA protein, human; D011944:Receptors, Androgen; D014105:Tosyl Compounds; D014157:Transcription Factors; D000077767:Panobinostat; C053541:bicalutamide