Weidong Xu

Long non-coding RNA metastasis associated in lung adenocarcinoma transcript 1 derived miniRNA as a novel plasma-based biomarker for diagnosing prostate cancer

Examining plasma RNA is an emerging non-invasive diagnosis technique. However, whether tumour-derived long non-coding RNAs (lncRNAs) in plasma can be used as a novel approach to detect human prostate cancer (PCa) has not yet been established. The study was divided into three parts: (1) the characteristics of PCa-related lncRNA fragments were systematically studied in the plasma or serum of 25 patients; (2) the source of the circulating lncRNA fragments was explored in vitro and in vivo; and (3) the diagnostic performance of metastasis associated in lung adenocarcinoma transcript 1 (MALAT-1) derived (MD) miniRNA was validated in an independent cohort of 192 patients. The expression levels of lncRNAs were measured by quantitative real time polymerase chain reaction (qRT-PCR). The MD-miniRNA copies were calculated using a standard curve in an area under the ROC curve (AUC)-receiver operating characteristic (ROC) analysis. Genome-wide profiling revealed that MALAT-1 and prostate cancer gene 3 (PCA3) are overexpressed in PCa tissues. Plasma lncRNAs probably exist in the form of fragments in a stable form. MD-miniRNA enters cell culture medium at measurable levels, and MD-miniRNA derived from human PCa xenografts actually enters the circulation in vivo and can be measured to distinguish xenografted mice from controls. In addition, plasma MD-miniRNA levels are significantly elevated in PCa patients compared to non-PCa patients (p<0.001). At a cut-off of 867.8 MD-miniRNA copies per microlitre of plasma, the sensitivity is 58.6%, 58.6% and 43.5% and the specificity is 84.8%, 84.8% and 81.6% for discriminating PCa from non-PCa, positive biopsy from negative biopsy and positive biopsy from negative biopsy, respectively. We conclude that MD-miniRNA can be used as a novel plasma-based biomarker for PCa detection and can improve diagnostic accuracy by predicting prostate biopsy outcomes. Further large-scale studies are needed to confirm our findings.

The prostate cancer-up-regulated long noncoding RNA PlncRNA-1 modulates apoptosis and proliferation through reciprocal regulation of androgen receptor

OBJECTIVE Emerging evidences implicate long noncoding RNAs (lncRNAs) are deregulated in cancer development. The purpose of the current study is to investigate the role of new lncRNA, named PlncRNA-1, in prostate cancer (CaP) pathogenesis. MATERIALS AND METHODS In this study, real-time q-PCR was used to demonstrate the expression of PlncRNA-1 in 16 pairs CaP tissues and matched normal tissues, 14 pairs CaP tissues and BPH tissues, 4 CaP cell lines, including LNCaP, LNCaP-AI, PC3, and C4-2, and 2 normal prostate epithelial cell lines RWPE-1 and PWR-1E. After PlncRNA-1 was suppressed by siRNA in LNCaP and LNCaP-AI cell lines, cell proliferation and apoptosis were assessed using CCK-8 and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). After PlncRNA-1 and AR was suppressed by siRNA in LNCaP and LNCaP-AI cell lines, real-time q-PCR and Western blotting were used to measure reciprocal regulation of PlncRNA-1 and AR. RESULTS We showed that expression PlncRNA-1, was significantly higher in CaP cells relative to normal prostate epithelial cells, as well as higher in human CaPs compared with normal tissues and benign prostatic hyperplasia (BPH). Silencing of PlncRNA-1 significantly reduced cell proliferation and induced apoptosis in CaP cell lines LNCaP and LNCaP-AI. Mechanistically, PlncRNA-1 suppression by siRNA resulted in a decrease of androgen receptor (AR) mRNA, protein and AR downstream target. Of note, blockade of AR signaling with siRNA also resulted in a suppression of PlncRNA-1 expression in CaP cell lines. CONCLUSIONS Our study suggests reciprocal regulation of PlncRNA-1 and androgen receptor contribute to CaP pathogenesis and that PlncRNA-1 is a potential therapy target.

miRNA-100 Inhibits Human Bladder Urothelial Carcinogenesis by Directly Targeting mTOR

miRNAs are involved in cancer development and progression, acting as tumor suppressors or oncogenes. In this study, miRNA profiling was conducted on 10 paired bladder cancer tissues using 20 GeneChip miRNA Array, and 10 differentially expressed miRNAs were identified in bladder cancer and adjacent noncancerous tissues of any disease stage/grade. After being validated on expanded cohort of 67 paired bladder cancer tissues and 10 human bladder cancer cell lines by quantitative real-time PCR (qRT-PCR), it was found that miR-100 was downregulated most significantly in cancer tissues. Ectopic restoration of miR-100 expression in bladder cancer cells suppressed cell proliferation and motility, induced cell-cycle arrest in vitro, and inhibited tumorigenesis in vivo both in subcutaneous and in intravesical passage. Bioinformatic analysis showed that the mTOR gene was a direct target of miR-100. siRNA-mediated mTOR knockdown phenocopied the effect of miR-100 in bladder cancer cell lines. In addition, the cancerous metastatic nude mouse model established on the basis of primary bladder cancer cell lines suggested that miR-100/mTOR regulated cell motility and was associated with tumor metastasis. Both mTOR and p70S6K (downstream messenger) presented higher expression levels in distant metastatic foci such as in liver and kidney metastases than in primary tumor. Taken together, miR-100 may act as a tumor suppressor in bladder cancer, and reintroduction of this mature miRNA into tumor tissue may prove to be a therapeutic strategy by reducing the expression of target genes. Mol Cancer Ther; 12(2); 207–19. ©2012 AACR.

Oncogenic CUL4A determines the response to thalidomide treatment in prostate cancer

Thalidomide is experimentally used to treat various human cancers; however, clinical responses to thalidomide are sporadic. Here we demonstrate that CUL4A plays an oncogenic role in prostate cancer development and prostate cancer cells with higher level of CUL4A are particularly sensitive to thalidomide treatment. We show that CUL4A is frequently overexpressed in human primary prostate cancer and cell lines. Notably, subjects with tumors that highly expressed CUL4A had poor overall survival. CUL4A downregulation inhibited cell proliferation and induced apoptosis in vitro and in vivo, whereas CUL4A overexpression transformed human normal prostate epithelial cells and promoted invasion, which was attenuated by the extracellular signal-regulated kinase (ERK) inhibitor. We further show that the sensitivity to thalidomide is positively correlated with CUL4A expression in a panel of prostate cell lines. Ectopic CUL4A expression greatly enhanced sensitivity to thalidomide, while its downregulation conferred resistance to this drug. Mechanistically, thalidomide decreased CUL4A in a time- and dose-dependent manner, consequently leading to inaction of ERK pathway. Finally, we show that cereblon level is correlated with CUL4A expression and downregulated in thalidomide-resistant prostate cancer cell. Our results offer the first evidence that CUL4A is a potential therapeutic target for prostate cancer and may serve as a biomarker for assessing prognosis of human prostate cancer and response to thalidomide treatment.

Histone methyltransferase SETDB1 is required for prostate cancer cell proliferation, migration and invasion

SETDB1 has been established as an oncogene in a number of human carcinomas. The present study was to evaluate the expression of SETDB1 in prostate cancer (PCa) tissues and cells and to preliminarily investigate the role of SETDB1 in prostate tumorigenesis in vitro. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) were used to detect the expression of SETDB1 in PCa tissues, adjacent normal tissues, benign prostatic hyperplasia (BPH) tissues, PCa cell lines and normal prostate epithelial cells. The results suggested that SETDB1 was upregulated in human PCa tissues compared with normal tissues at the mRNA and protein levels. The role of SETDB1 in proliferation was analyzed with cell counting kit-8, colony-forming efficiency and flow cytometry assays. The results indicated that downregulation of SETDB1 by siRNA inhibited PCa cell growth, and induced G0/G1 cell cycle arrest. The PCa cell migration and invasion decreased by silcencing SETDB1 which were assessed by using in vitro scratch and transwell invasion assay respectively. Our data suggested that SETDB1 is overexpressed in human PCa. Silencing SETDB1 inhibited PCa cell proliferation, migration and invasion.

Utility of a Modality Combining FISH and Cytology in Upper Tract Urothelial Carcinoma Detection in Voided Urine Samples of Chinese Patients

OBJECTIVES To evaluate a combined analysis approach that involves cytologic evaluation and fluorescence in situ hybridization analysis (FISH) for detecting urothelial carcinoma (UC) in the upper tract (UT). METHODS By refining the UroVysion positive criteria, an analyzing modality (Cyto-FISH) combined urine cytology and FISH analysis (UroVysion probe set) was introduced and urine specimens from 71 patients with UT-UC and 45 controls were analyzed. Sensitivity and specificity of urine cytology, FISH, and Cyto-FISH were determined and compared, respectively. The features of chromosomal aberrations of malignant cells from UT-UC were also determined. RESULTS Overall sensitivity of verified UT-UC by Cyto-FISH analysis was sharply higher than the single value for urine cytology (85.9% vs 45.1%, P <.001) and was slightly higher than FISH (85.9% vs 78.9%, P = .378). Sensitivities of cytology and Cyto-FISH by grade were 28.2% vs 74.4% for low-grade (P <.001), and 65.6% vs 96.9% for high-grade tumors (P = .003), respectively. The advantage maintains stably not only in the detection of nonmuscle-invasive tumors but in invasive tumors between cytology and Cyto-FISH (39.1% vs 76.1%, P = .001, and 53.8% vs 100%, P <.001, respectively). Specificities were 97.8%. In addition, polysomic chromosomal aberrations of the UT-UC cases could present a possible trend toward greater chromosome increased with tumor grades and progressive stages of invasion. CONCLUSIONS Our results have shown that Cyto-FISH analysis for the presence of UC cells is a powerful tool, providing high sensitivity and specificity, and may offer a new scheme for the tough UT-UC diagnosis.

Relationship between serum sex hormones levels and degree of benign prostate hyperplasia in Chinese aging men

Benign prostatic hyperplasia (BPH) is one of the most common medical conditions in middle aged and older men. This study investigated the relationship between serum levels of sex hormones and measures of BPH in the aging male population of China. Prostate symptoms were assessed as part of a free health screening program for men ≥ 40 years of age. The examination included digital rectal examination, determination of serum prostate-specific antigen levels, International Prostate Symptom Score (IPSS) and transrectal ultrasonography. Serum levels of total testosterone (TT), sex hormone binding globulin (SHBG), free testosterone (FT), luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin (PRL) and estradiol (E(2)) were evaluated. The men also completed a health and demographics questionnaire and received a detailed physical examination. The final study population consisted of 949 men with a mean age of 58.9 years. Pearson correlation analysis indicated that there were significant correlations between age and levels of all sex hormones except TT, and between age and prostate volume (PV; r=0.243; P<0.01) or IPSS (r=0.263; P<0.01). Additional significant correlations were found between IPSS and serum levels of LH (r=0.112; P<0.01) and FSH (r=0.074; P<0.05), but there were no significant correlations between sex hormone levels and PV. Multivariate linear regression analysis showed significant correlations between age and body mass index (BMI) with PV (P<0.0001). In addition, there was a significant correlation between age and PV with IPSS (P<0.0001). Serum sex hormone levels did not correlate with PV or IPSS. The effects of endocrine changes on measures of BPH in aging men require further investigation in longitudinal and multicenter studies that include patients with all severities of BPH.

A Pilot Study of Vela Laser for En Bloc Resection of Papillary Bladder Cancer

Micro‐Abstract Transurethral resection of bladder tumors is associated with perioperative or postoperative complications, and an “incise and scatter” procedure contradicts basic oncologic principles. The present study introduces the Vela laser, a new type of thulium laser with a 1.94‐&mgr;m wavelength, for en bloc resection of bladder tumors. The pilot experience found the Vela laser could preserve the muscle layer and was effective, feasible, and safe for patients with bladder tumors. Introduction: The present study evaluated the safety and efficacy of the Vela laser for en bloc resection of papillary bladder tumors. Materials and Methods: From January 2013 to August 2014, 38 patients underwent en bloc resection with the Vela laser and a 26F continuous flow resectoscope or 18F flexible cystoscope. Random cold forceps biopsy samples were also taken. The total operation time, pathologic result, and intraoperative and postoperative complications were recorded. Each patient was followed up for ≥ 1 year. Results: The average total operation time was 23 minutes. The en bloc resection of all tumors was successful, with 2 cases located at the bladder dome requiring the use of a flexible cystoscope for better management. No complications occurred during or after surgery. All resected tumors were intact with the detrusor muscle layer and architecture available for pathologic evaluation. One patient with stage T2b tumor underwent laparoscopic cystectomy 1 week after the initial surgery. At a median follow‐up period of 21.8 months, the recurrence rate at 12 months was 21.6% (8 of 37). Conclusion: The results of our study have shown that the Vela laser is an effective, feasible, and safe thulium laser for en bloc bladder tumor resection. It was associated with negligible complications and allows accurate pathologic evaluation. The Vela laser can serve as an alternative treatment method for nonmuscle‐invasive bladder cancer or infiltrating tumor.

Whole-exome sequencing of muscle-invasive bladder cancer identifies recurrent copy number variation in IPO11 and prognostic significance of importin-11 overexpression on poor survival

Non-muscle-invasive bladder cancer (NMIBC) often has a worse prognosis following its progression to muscle-invasive bladder cancer (MIBC), despite radical cystectomy with pelvic lymph node dissection combined with chemotherapy. Therefore, the discovery of novel biomarkers for predicting the progression of this disease and of therapeutic targets for preventing it is crucial. We performed whole-exome sequencing to analyze superficial tumor tissues (Tsup) and basal tumor tissues (Tbas) from 3 MIBC patients and identified previously unreported copy number variations in IPO11 that warrants further investigation as a molecular target. In addition, we identified a significant association between the absolute copy number and mRNA expression of IPO11 and found that high importin-11 expression was correlated with poor 3-year overall survival (OS), cancer-specific survival (CSS) and cancer-free survival (CFS) compared with low expression in the BCa patients. Importin-11 overexpression was also an independent risk factor for CSS and CFS in the BCa patients. Our study has revealed that IPO11 copy number amplification contributes to its overexpression and that these changes are unfavorable prognostic factors in NMIBC. Thus, IPO11 copy number amplification and importin-11 overexpression are promising biomarkers for predicting the progression and poor prognosis of patients with NMIBC.

Imaging diagnosis and endoscopic treatment for ureteral fibroepithelial polyp prolapsing into the bladder

OBJECTIVE Fibroepithelial polyps of ureter prolapsing into the bladder are a rare urological condition. We report the imaging findings and our experience with endoscopic treatment for ureteral fibroepithelial polyps prolapsing into the bladder. PATIENTS AND RESULTS Four patients with frank pain and hematuria were enrolled. Intravenous urography and computed tomography revealed a ureteral mass with filling defects in affected ureter and mild hydronephrosis. Endoscopic examination showed ureteral polyps prolapsing in the bladder. The histopathologic diagnosis on 4 cases was benign fibroepithelial polyps of ureter. The largest polyps (from 4-10 cm in length) were successfully resected and vaporized by Holmium: YAG laser. A double-pigtail ureteral stent at 7F was placed and left for 6 weeks after the procedure. Neither recurrence nor ureter stricture was observed after up to 12 years of follow-up. CONCLUSIONS Ureteral malignancy must be excluded in cases where a ureteral mass is detected. Endoscopic management is recommended to minimize morbidity and complications in treatment of ureteral fibroepithelial polyps that prolapse into the bladder.