Identification of miRs-143 and -145 that Is Associated with Bone Metastasis of Prostate Cancer and Involved in the Regulation of EMT

The principal problem arising from prostate cancer (PCa) is its propensity to metastasize to bone. MicroRNAs (miRNAs) play a crucial role in many tumor metastases. The importance of miRNAs in bone metastasis of PCa has not been elucidated to date. We investigated whether the expression of certain miRNAs was associated with bone metastasis of PCa. We examined the miRNA expression profiles of 6 primary and 7 bone metastatic PCa samples by miRNA microarray analysis. The expression of 5 miRNAs significantly decreased in bone metastasis compared with primary PCa, including miRs-508-5p, -145, -143, -33a and -100. We further examined other samples of 16 primary PCa and 13 bone metastases using real-time PCR analysis. The expressions of miRs-143 and -145 were verified to down-regulate significantly in metastasis samples. By investigating relationship of the levels of miRs-143 and -145 with clinicopathological features of PCa patients, we found down-regulations of miRs-143 and -145 were negatively correlated to bone metastasis, the Gleason score and level of free PSA in primary PCa. Over-expression miR-143 and -145 by retrovirus transfection reduced the ability of migration and invasion in vitro, and tumor development and bone invasion in vivo of PC-3 cells, a human PCa cell line originated from a bone metastatic PCa specimen. Their upregulation also increased E-cadherin expression and reduced fibronectin expression of PC-3 cells which revealed a less invasive morphologic phenotype. These findings indicate that miRs-143 and -145 are associated with bone metastasis of PCa and suggest that they may play important roles in the bone metastasis and be involved in the regulation of EMT Both of them may also be clinically used as novel biomarkers in discriminating different stages of human PCa and predicting bone metastasis

Erectile Dysfunction in Chronic Prostatitis/Chronic Pelvic Pain Syndrome: Outcomes from a Multi-Center Study and Risk Factor Analysis in a Single Center.

The aim of this study was to investigate the prevalence of erectile dysfunction (ED) in patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) and explore the influence of UPOINT domains, National Institutes of Health-CP symptom index (NIH-CPSI) and other factors on ED prevalence. This was a prospective study of consecutive patients with CP/CPPS seen at 11 tertiary hospitals during January-July 2014. ED was diagnosed as a score of<21 on the International Index of Erectile Function (IIEF-5). Patients from one center were evaluated by the UPOINT system and NIH-CPSI. Each patient was assessed using clinical examination, asocio-demographic questionnaire, the Patient Health Questionnaire (PHQ), the Pain Catastrophizing Scale (PCS), NIH-CPSI and IIEF-5.1406 patients from 11 centers (mean age, 32.18 years; range 18-60 years) were enrolled. ED was found in 638/1406 patients (45.4%), and was categorized as mild in 291(45.6%), moderate in 297(46.6%) and severe in50(7.7%). 192 patients from one center(mean age,31.3 years; range 18-57 years) were further studied.IIEF-5 score correlated negatively with NIH-CPSI(r = 0.251), PHQ (r = 0.355) and PCS (r = 0.322)scores (P<0.001).PHQ score correlated positively with NIH-CPSI (r = 0.586) and PCS(r = 0.662) scores (P<0.001).NIH-CPSI, PHQ, PCS and IIEF-5 scores did not differ significantly between class IIIA and IIIB CP/CPPS. Multivariate logistic regression showed that UPOINT psychological (P) domain and NIH-CPSI symptom severity were independent risk factors for ED in CP/CPPS. It is concluded that psychological factors and symptom severity are independent risk factors for ED in CP/CPPS

Comparison of NIH-CPSI, PHQ, PCS and IIEF-5 scores between patients with type IIIA and IIIB CP/CPPS.

<p>Comparison of NIH-CPSI, PHQ, PCS and IIEF-5 scores between patients with type IIIA and IIIB CP/CPPS.</p

Multivariate logistic regression analysis of the influence of UPOINT domains and other factors on ED prevalence in men with CP/CPPS.

<p>Multivariate logistic regression analysis of the influence of UPOINT domains and other factors on ED prevalence in men with CP/CPPS.</p

Study flowchart for patients enrollment.

<p>Study flowchart for patients enrollment.</p

Differentially expressed miRNAs identified in bone metastasis of prostate cancer compared to primary prostate cancer by miRNA microarray.

<p>*Significantly differentially expressed miRNAs selected as following standards: |Score(d)|≥2, Fold Change≥2 or ≤0.5, q-value(%)≤5.</p

Both of miRs-143 and -145 repressed the development and invasion of PC-3 cells in bone.

<p>Male SCID mice were inoculated with PC-3 cells through the intra-tibial route. Skeletal lesions in radiographs are demonstrated by arrows (upper panel), and histologic analysis was carried by H&E-staining in which tumors were lined out by dashed line and marked as “T” (middle panel, taken under microscope 40×). Lesion scores of control and experimental specimens were shown in lower panels, where results were showed by means ± SEM of each group, <i>p</i> = 0.035 and <i>p</i> = 0.014 respectively.</p

Validation of select miRNAs predicted to be downregulated in prostate cancer.

<p><i>A,</i> The certified result of microarray analysis. <i>B,</i> Real-time RT–PCR assays on miR-143 (left panel), miR-145 (right panel), and miR-125b (bottom panel) in 16 primary prostate cancer and 13 bone metastasis tissues. The order of the tissue samples is the same for all three plots. <i>p</i>-values by <i>t-test</i>.</p

Detection of miRs-143 and -145 in primary PCa tissues by ISH.

<p>Sections at upper panel showed the identification of tumor cells and stromal cells by H&E-stainning. Sections at lower panel showed the location of miR-143 (left) and miR-145 (right) in PCa cells by LNA-ISH. Signals of miRs-143 and -145 were in purple blue. Pictures were taken under microscope 200×.</p

Construction of stable cell lines.

<p><i>A–B,</i> Ectopic expression of miRs-143 or -145 in PC-3 (A) and LNCaP (B) were verified by qRT-PCR (<i>t-test</i>, <i>p</i><0.01).</p