Effects of the dual 5 α‐reductase inhibitor dutasteride on apoptosis in primary cultures of prostate cancer epithelial cells and cell lines

Abstract

BACKGROUNDThe profound reduction in serum dihydrotestosterone (DHT) observed with the dual 5 α‐reductase inhibitor (5ARI) dutasteride makes it an attractive agent for prostate cancer therapy. The objective of the current study was to determine whether dutasteride would induce apoptosis in a range of prostate epithelial cell lines and primary cultures.METHODSBoth human prostate androgen‐sensitive cell lines (PwR‐1E, PNT‐2, LNCaP, and PC3[AR2]) and an androgen‐independent cell line (PC‐3) were grown to confluence. Primary epithelial cells extracted from fresh prostate cancer radical prostatectomy specimens also were grown to confluence under optimal conditions. Total cellular protein was extracted to confirm cytokeratin 18 and antihuman α‐methylacyl‐CoA racemase (AMACR) expression of the primary cells. Apoptosis was assessed by propidium iodide DNA staining and flow cytometry after 24 hours of culture in from 0 μM to 10 μM of dutasteride.RESULTSDutasteride induced a dose‐dependent increase in apoptosis in the androgen‐sensitive prostate cell lines PwR‐1E, PNT‐2, and LNCaP and in the androgen receptor‐expressing PC3(AR2) cell line. However, there was no significant apoptosis noted in the parental PC‐3 cells. Of 16 primary epithelial cultures that were treated, 7 cultures were induced to undergo apoptosis, and 9 cultures were unresponsive. All primary cultures were positive for cytokeratin 18 expression, confirming their epithelial phenotype. Responder epithelial cells were positive for AMACR expression.CONCLUSIONSThe results of the current study confirmed that dutasteride differentially induced apoptosis in a subset of prostate cell lines and primary prostate epithelial cells. Understanding the cellular phenotype may indicate susceptible cells. Cancer 2006. © 2006 American Cancer Society.

Concentration‐dependent effects of dutasteride on prostate‐specific membrane antigen (PSMA) expression and uptake of 177Lu‐PSMA‐617 in LNCaP cells

“…Since dutasteride is widely used among the male population for the treatment benign prostatic enlargement, it could easily be implemented into clinics as a possible PSMA expression enhancer before PSMA‐based imaging or as synergistic PSMA expression enhancer before 177 Lu‐PSMA‐617 radioligand therapy . All dutasteride concentrations tested in our in vitro studies have previously been used by others in different settings . However, the highest serum drug concentrations observed in humans reached above 1 μM in men given an oral dose of 5 mg dutasteride daily .…”

“…12 All dutasteride concentrations tested in our in vitro studies have previously been used by others in different settings. [22][23][24][25] However, the highest serum drug concentrations observed in humans reached above 1 μM in men given an oral dose of 5 mg dutasteride daily. 26 Therefore, most concentrations of dutasteride used for in vitro and in vivo experiments are significantly higher than serum levels of dutasteride observed in clinical trials.…”

Characterisation and manipulation of docetaxel resistant prostate cancer cell lines

“…Total RNA was extracted using Trizol reagent (Invitrogen, Paisley, UK) using standard procedures as previously described [17]. The RNA was used to generate cDNA as previously described [17].…”

“…The RNA was used to generate cDNA as previously described [17]. PCR amplification of cDNA template was performed in a thermal cycler (Perkin Elmer 7700).…”

Cancer Biomarker Discovery: The Entropic Hallmark

“…Here we present results, from an unifying biological and informational principle, which allows (using Ref. [332]'s own data) the identification of the most central current biomarker with a truly compelling body of support in independent studies [316], [340], [341], [342], [343], [344], [345], [346], [347], [348], [349], [350], [351], [352], [353], [354], [355], [356], [357], [358], [359], [360], [361], [362], [363], [364], [365], [366], [367], [368], [369], [370], [371], [372], [373], [374], [375], [376], [377], [378], [379], [380], [381], [382], [383], [384], [385], [386], [387], [388], [389], [390], [391], [392], [393], [394], [395], [396], [397], [398], [399], …”