A novel platform using homobifunctional hydrazide for enrichment and isolation of urinary circulating RNAs

Abstract Changes in specific circulating RNA (circRNA) expressions can serve as diagnostic noninvasive biomarkers for prostate cancer (PCa). However, there are still unmet needs, such as unclear types and roles of circRNAs, PCa detection in benign prostatic hyperplasia (BPH) by unstandardized methods, and limitations of sample volume capacity and low circRNA concentrations. This study reports a simple and rapid circRNA enrichment and isolation technique named “HAZIS‐CirR” for the analysis of urinary circRNAs. The method utilizes homobifunctional hydrazides with amine‐modified zeolite and polyvinylidene fluoride (PVDF) syringe filtration for combining electrostatic and covalent coupling and size‐based filtration, and it offers instrument‐free isolation of circRNAs in 20 min without volume limitation, thermoregulation, and lysis. HAZIS‐CirR has high capture efficiency (82.03%–92.38%) and a 10‐fold more sensitive detection limit (20 fM) than before enrichment (200 fM). The clinical utility of HAZIS‐CirR is confirmed by analyzing circulating mRNAs and circulating miRNAs in 89 urine samples. Furthermore, three miRNA panels that differentiate PCa from BPH and control, PCa from control, and BPH from control, respectively, are established by comparing miRNA levels. HAZIS‐CirR will be used as an optimal and established method for the enrichment and isolation of circRNAs as diagnostic, prognostic, and predictive biomarkers in human cancers

DataSheet_1_Glucocorticoid receptor and androgen receptor-targeting therapy in patients with castration-resistant prostate cancer.docx

ObjectiveThe glucocorticoid receptor (GR) promotes resistance to androgen receptor (AR)-targeting therapies in castration-resistant prostate cancer (CRPC) by bypassing AR blockade. However, the clinical relevance of evaluating GR expression in patients with CRPC has not been determined. The present study investigated the association of relative GR expression in CRPC tissue samples with treatment response to AR-targeting therapy.MethodsLevels of GR, AR-FL, and AR-V7 mRNAs were measured in prostate cancer tissue from prospectively enrolled CRPC patients who were starting treatment. Patients were divided into groups with high and low AR-V7/AR-FL ratios and with high and low GR/AR-FL ratios. The primary endpoint was prostate-specific antigen (PSA) response rate to treatment.ResultsEvaluation of 38 patients treated with AR-targeting therapies showed that the PSA response rate was significantly higher in patients with low than high AR-V7/AR-FL ratios (77.8% vs. 25.0%, p=0.003) and in patients with low than high GR/AR-FL ratios (81.3% vs. 27.3%, p=0.003). Patients with low GR/AR-FL ratios had higher rates of PSA progression-free survival (46.0% vs. 22.4%, p=0.006), radiologic progression-free survival (28.9% vs. 10.0%, p=0.02), and overall survival (75.2% vs. 48.0%, p=0.037) than patients with high GR/AR-FL ratios. The association of GR/AR-FL ratio with PSA response to AR-targeting therapy remained significant in multivariable models. Evaluation of the 14 patients who received taxane chemotherapy showed that PSA response rates did not differ significantly in those with low and high AR-V7/AR-FL and GR/AR-FL ratios, although no definitive conclusions can be drawn due to the small number of patients.ConclusionRelative GR expression is associated with sensitivity to AR-targeting therapy and survival in patients with CRPC. Large-scale prospective validation and liquid biopsy-based studies are warranted.</p

Correction: KML001 Induces Apoptosis and Autophagic Cell Death in Prostate Cancer Cells via Oxidative Stress Pathway.

[This corrects the article DOI: 10.1371/journal.pone.0137589.]

Structural features observed by electron microscope (10000× and 5000×) in PC3, DU145, and LNCaP prostate cancer cells treated with KML001 for 24 and 48 h.

<p>Structural features observed by electron microscope (10000× and 5000×) in PC3, DU145, and LNCaP prostate cancer cells treated with KML001 for 24 and 48 h.</p

KML001 treatment has (A) anti-proliferative effect, (B) apoptotic effect, and (C) autophagic effect on DU145 prostate cancer cells in mice.

<p>Three fields in each mice were observed by fluorescence (200×) and bright field microscopes (200×), respectively. Representative images of each treatment group were shown. Vehicle and KML001 group mice orally received saline and KML001 (2.5 or 10 mg/kg/d) for 4 weeks, respectively. * <i>p</i> < 0.05 vs. vehicle.</p

IC₅₀ values of KML001 in prostate cancer cells.

<p>IC₅₀ values of KML001 in prostate cancer cells.</p

Growth inhibition of KML001 in prostate cell lines.

<p>Cells treated with various concentrations of KML001 were incubated for 72 h. The Alamar blue assay was done in triplicate. Mean values are given, the value of control being 100%. PC3 (●, <i>thick solid line</i>), DU145 (▲, <i>thin solid line</i>), and LNCaP (▼, <i>thin solid line</i>).</p