Journal Article

Growth inhibition of androgen-responsive prostate cancer cells with brefeldin A targeting cell cycle and androgen receptor

Choudhury, Muhammad, Rajamahanty, Srinivas, Aynehchi, Shahrad, Konno, Sensuke, Alonzo, Catherine

2010

Overview

Background Androgen ablation is one of the viable therapeutic options for patients with primary hormone (androgen)-dependent prostate cancer. However, an antibiotic brefeldin A (BFA) has been shown to exhibit the growth inhibitory effect on human cancer cells. We thus investigated if BFA might inhibit proliferation of androgen-responsive prostate cancer LNCaP cells and also explored how it would be carried out, focusing on cell cycle and androgen receptor (AR). Methods Androgen-mediated cellular events in LNCaP cells were induced using 5α-dihydrotestosterone (DHT) as an androgenic mediator. Effects of BFA on non-DHT-stimulated or DHT-stimulated cell growth were assessed. Its growth inhibitory mechanism(s) was further explored; performing cell cycle analysis on a flow cytometer, assessing AR activity by AR binding assay, and analyzing AR protein expression using Western blot analysis. Results DHT (1 nM) was capable of stimulating LNCaP cell growth by ~40% greater than non-stimulated controls, whereas BFA (30 ng/ml) completely inhibited such DHT-stimulated proliferation. Cell cycle analysis showed that this BFA-induced growth inhibition was associated with a ~75% reduction in the cell number in the S phase and a concomitant increase in the G 1 cell number, indicating a G 1 cell cycle arrest. This was further confirmed by the modulations of specific cell cycle regulators (CDK2, CDK4, cyclin D 1 , and p21 WAF1 ), revealed by Western blots. In addition, the growth inhibition induced by BFA was accompanied by a profound (~90%) loss in AR activity, which would be presumably attributed to the significantly reduced cellular AR protein level. Conclusions This study demonstrates that BFA has a potent growth inhibitory activity, capable of completely inhibiting DHT (androgen)-stimulated LNCaP proliferation. Such inhibitory action of BFA appears to target cell cycle and AR: BFA led to a G 1 cell cycle arrest and the down-regulation of AR activity/expression, possibly accounting for its primary growth inhibitory mechanism. Thus, it is conceivable that BFA may provide a more effective therapeutic modality for patients with hormone-dependent prostate cancer.

• Publisher: BioMed Central,BioMed Central Ltd,BMC
• Subject: Analysis / Androgens / Antineoplastic Agents - pharmacology / Biomedical and Life Sciences / Biomedicine / Brefeldin A - pharmacology / Cancer / Cancer cells / Care and treatment / Cell cycle / Cell Cycle - drug effects / Cyclin D - metabolism / Cyclin-Dependent Kinase 2 - metabolism / Cyclin-Dependent Kinase 4 - metabolism / Dihydrotestosterone - pharmacology / G1 Phase - drug effects / Growth / Growth Inhibitors - pharmacology / Health aspects / Humans / Male / Prevention / Prostate cancer / Prostatic Neoplasms - metabolism / Protein binding / Receptors, Androgen - genetics / Receptors, Androgen - metabolism / Tumor Cells, Cultured