Survival analysis of patients with invasive extramammary Paget disease: implications of anatomic sites.

BACKGROUND: Extramammary Paget disease (EMPD) is a rare malignant dermatosis with poorly defined outcomes. We investigated clinical characteristics of invasive EMPD at different anatomic sites and by subject demographics to determine prognostic factors for overall survival (OS). METHODS: All patient data were collected from the Surveillance, Epidemiology, and End Results (SEER) program, 1973-2013, of the U.S. National Cancer Institute. Patients with invasive EMPD of skin, vulva/labia, vagina, scrotum/penis, or other sites were included. After excluding patients with unknown radiation status, data of 2001 patients were analyzed. Primary endpoint was EMPD mortality by anatomic sites. Independent variables included patients' demographic data, concurrent malignancy (ie, non-EMPD related cancers), tumor size, distant metastasis, and surgery and/or radiation or not. RESULTS: Multivariate regression analysis showed that mortality was significantly higher in patients with vaginal EMPD than in patients with vulvar/labial EMPD (adjusted hazard ratio [aHR] = 3.26, p < 0.001). Patients with distant metastasis had higher mortality than those without (aHR = 3.36, p < 0.001). Patients who received surgery had significantly lower mortality than those who did not receive surgery (aHR = 0.77, p = 0.030), and those treated with radiation had significantly higher mortality than those who did not receive radiation (aHR = 1.60, p = 0.002). Older age was associated with significantly increased mortality (aHR = 1.09, p < 0.001), and mortality was significantly higher in males than in females (aHR = 1.42, p = 0.008). CONCLUSIONS: In conclusion, among EMPD patients, mortality is higher in patients with vaginal EMPD than in those with vulvar/labial EMPD and higher in those who are older, those with concurrent malignancy or distant metastasis. Mortality is also higher in males than in females. Surgery is a protective factor and radiation is a risk factor for OS. Greater understanding of EMPD clinical characteristics, and considering EMPD in differential diagnosis of chronic genital and perianal dermatoses may provide support for early EMPD diagnosis and definitive surgical treatment.

["3+1" bladder function restoration combined with holmium laser enucleation of the prostate for benign prostatic hyperplasia with acontractile detrusor].

OBJECTIVE: To investigate the clinical effect of "3+1" bladder function restoration combined with holmium laser enucleation of the prostate (HoLEP) in the treatment of benign prostatic hyperplasia (BPH) with acontractile detrusor (ACD). METHODS: We treated 35 BPH patients with ACD by HoLEP followed by "3+1" bladder function restoration, that is, a 3-phase bladder function training plus simultaneous 1-drug medication after surgery. We recorded and analyzed the detrusor pressure, post-void residual urine volume (PVR), maximum urinary flow rate (Qmax), International Prognostic Scoring System (IPSS) scores, quality of life (QoL), voluntary micturition, satisfaction with the bladder function, hydronephrosis, ureterectasia, renal function, and urinary tract infection of the patients before and after treatment. RESULTS: Compared with the base line, at 6 months treatment, the patients showed significantly increased detrusor pressure (ï¼»35.1±2.7ï¼½vs ï¼»50.2±2.3ï¼½ cmH2O, P<0.05) and Qmax (ï¼»4.2±2.7ï¼½vs ï¼»21.1±4.1ï¼½ ml/s, P<0.05) but decreases in PVR (ï¼»173.0±31.6ï¼½ vs ï¼»30.5±12.9ï¼½ml, IPSS score (27.3±3.2 vs 5.1±1.4, P<0.05) and QoL (4.1±0.8 vs 0.8±0.1, P<0.05), elevated rates of voluntary urination (0% ï¼»0/35ï¼½ vs 100% ï¼»35/35ï¼½, P<0.05), regularurination (0% ï¼»0/35ï¼½ vs 85.71% ï¼»30/35ï¼½, P<0.05), grade Ⅰ satisfaction with bladder function (0% ï¼»0/35ï¼½ vs 85.71% ï¼»30/35ï¼½, P<0.05), reduced rate of overflowing urinary incontinence (28.57% ï¼»10/35ï¼½ vs 5.71% ï¼»2/35ï¼½, P<0.05), and increased percentages of normal renal function (34.29% ï¼»12/35ï¼½ vs 85.71% ï¼»30/35ï¼½, P<0.05) and non-infection of the urinary system (17.14% ï¼»6/35ï¼½ vs 94.29% ï¼»33/35ï¼½, P<0.05). After treatment, urination was markedly improved in 94.29% (33/35) of the patients. CONCLUSIONS: "3+1" bladder function restoration combined with HoLEP produced a desirable effect on BPH with ACD, though its long-term effect remains to be further investigated.

Traditional Chinese herbal medicine retention enema combined with perineal massage (THREM): a promising therapeutic strategy for refractory chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS).

Background: Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a debilitating condition characterized by lower urinary tract symptoms and persistent pelvic pain or discomfort lasting for more than three months. Currently available oral drug therapies exhibit limited efficacy in the treatment of CP/CPPS. Therefore, personalized and combination therapies are recommended by Chinese CP/CPPS guidelines, which primarily include traditional Chinese medicine, radiofrequency therapy, urethral lavage, transrectal prostate massage, extracorporeal shock wave therapy. However, a significant number of patients do not respond well to all types of these therapeutic methods. Among those who have sequentially or simultaneously undergone at least three different treatment modalities, in addition to oral medications, for more than 1 year, they are defined as patients with refractory CP/CPPS. This retrospective study aims to evaluate the clinical effect of traditional Chinese herbal medicine retention enema combined with perineal massage (THREM) in managing refractory CP/CPPS. Methods: A total of 20 patients with refractory CP/CPPS, who did not show significant improvement despite receiving multiple conventional treatments, including oral medications, were included in this study. Following THREM therapy, the International Prostate Symptom Score (IPSS), visual analogue scale (VAS), and National Institutes of Health-Chronic Prostatitis Symptom Index (NIH-CPSI) quality of life (QoL) score were used to assess treatment efficacy. Results: Six months after THREM therapy, a significant decrease in IPSS, VAS, and QoL scores was observed (P<0.01). Importantly, 85% of the patients experienced a reduction in symptoms of ≥60%, with an average degree of alleviation reaching 70.25%±24.20%. Conclusions: THREM treatment demonstrated excellent efficacy in managing refractory CP/CPPS at least for 6 months. It has promising clinical application prospects. Further research is warranted to validate these results and explore the underlying mechanisms of THREM therapy.

A prospective, randomized clinical trial comparing plasmakinetic resection of the prostate with holmium laser enucleation of the prostate based on a 2-year followup.

PURPOSE: We compared plasmakinetic resection with holmium laser enucleation of the prostate for the treatment of benign prostatic hyperplasia by analyzing 2-year followup data from a prospective randomized clinical trial. MATERIALS AND METHODS: A total of 280 patients were randomly treated with plasmakinetic resection or holmium laser enucleation of the prostate. Perioperative and postoperative outcome data were obtained during a 2-year followup. RESULTS: No significant differences between the 2 surgical groups were observed in the preoperative data. Both groups displayed significant improvements after surgery. However, we identified no significant differences between the 2 groups in the 2-year followup data for I-PSS (International Prostate Symptom Score), quality of life scores or maximum flow rate values. Patients in the holmium laser enucleation group displayed a lower risk of hemorrhage, shorter bladder irrigation and catheter times, and shorter hospital stays. A larger amount of prostate tissue was retrieved in the holmium laser enucleation group, but the operation time was longer for this group than for the plasmakinetic resection group. CONCLUSIONS: Plasmakinetic resection and holmium laser enucleation of the prostate are effective and safe treatments for benign prostatic hyperplasia. Holmium laser enucleation of the prostate can be applied to prostates of all sizes, and involves less risk of hemorrhage, decreased bladder irrigation and catheter times, as well as reduced hospital stay. Thus, we believe holmium laser enucleation of the prostate should be proposed as a potential new gold standard surgical therapy instead of transurethral resection of the prostate for patients with benign prostatic hyperplasia.

Poly(ADP-ribose) polymerase inhibition improves erectile function by activation of nitric oxide/cyclic guanosine monophosphate pathway in diabetic rats.

INTRODUCTION: Endothelial dysfunction-induced abnormalities of the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) signaling pathway in the corpus cavernosum are thought to be the main factors involved in the pathogenesis of diabetes-induced erectile dysfunction (ED). Recent studies have shown that the poly(adenosine diphosphate ribose) polymerase (PARP) pathway plays a critical role in diabetic endothelial dysfunction. AIM: The aim of this study is to determine whether activation of the PARP pathway is involved in diabetic cavernosal endothelial dysfunction and abnormalities of the NO/cGMP pathway. METHODS: Male Sprague-Dawley rats were randomly divided into three groups: age-matched controls, diabetic controls (DM), and the 3-aminobenzamide (3-AB, a PARP inhibitor)-treated diabetic group (DM+3-AB). Diabetes was induced by intraperitoneal injection of streptozotocin. Eight weeks after inducing diabetes, the DM+3-AB group was treated with 3-AB for 4 weeks. MAIN OUTCOME MEASURES: Erectile function was assessed at 12 weeks after inducing diabetes by stimulating the cavernous nerve. Expression of poly(ADP-ribose), protein kinase B (Akt), phospho-Akt, endothelial nitric oxide synthase (eNOS), phospho-eNOS, and neuronal nitric oxide synthase (nNOS) were evaluated by Western blot. Cavernous NO generation and cGMP levels were also determined. RESULTS: The DM group showed impaired erectile function and significantly increased PARP activity. Expression of total eNOS and nNOS, phospho-Akt, and eNOS decreased significantly in the DM group compared with those in the control group. In addition, cavernous NO generation and cGMP levels decreased significantly in the DM group compared with those in the control group. Treatment with 3-AB restored erectile function and significantly reversed all molecular alterations except decreased nNOS expression. CONCLUSION: Overactivation of the PARP pathway in the corpus cavernosum of diabetic rats was involved in cavernosal endothelial dysfunction and abnormalities of the NO/cGMP pathway resulting in ED. These findings may be applied to develop novel therapies for patients with diabetic ED.

NLRP3+ macrophages aggravate inflammatory cystitis in diabetes.

Inflammatory macrophages play a pivotal role in the progression of inflammatory cystitis. Formation of NOD-, LRR- and PYD domains-containing protein 3 (NLRP3) inflammasome triggers the activation of caspase-1/IL-1ß signaling cascades to mediate inflammatory response. However, it is not known whether NLRP3 activation in macrophages during cystitis may differ in normal or diabetic setting as well as the importance of it. In this study, we found that NLRP3 levels significantly increased in bladder macrophages in diabetic mice that underwent cystitis. Moreover, bladder macrophages from diabetic mice appeared to have increased their potential of growth, migration and phagocytosis. Furthermore, specific depletion of NLRP3 in macrophages alleviated the severity of cystitis in diabetic mice, but not in non-diabetic mice. Together, our data suggest that NLRP3 depletion in macrophages may be a promising strategy for treating diabetic cystitis.

Loss of NLRP3 increases bacterial cystitis via IRAKM.

Background: We attempted to characterize the molecular mechanisms that underpin urinary tract infections using a mouse model of cystitis induced by bacterial infection in a background of NOD-, LRR- and PYD domains-containing protein (NLRP3) deficiency. Methods: Male NLRP3 knockout (NLRP3-/-) and control mice (12 weeks old) were intraurethrally inoculated with 2×108 Escherichia coli (E. coli) and euthanized 1, 3, and 7 days later to assess the degree of bladder infection. Immunohistochemical detection of NLRP3 and interleukin-1 receptor-associated kinase M (IRAKM) was performed. Quantitative PCR analysis was performed to analyze the expression of interleukin (IL)-1ß and tumor necrosis factor (TNF)-α. Results: Bladder infection was observed in control mice 1 day after inoculation with E. coli. The infection had disappeared by day 7. IL-1ß and TNF-α levels were lower 1 day after injection but higher on days 3 and 7 in the NLRP3-/- group compared with the control mice (P<0.05). Expression of NLRP3 and IRAKM in wild-type (WT) group were significantly decreased 1 day post infection, and by day 7 were increased back to similar level on day 0. On the contrary, in the NLRP3-/- group, IRAKM was significantly lower than WT mice on day 0 and were significantly decreased by day 7. Conclusions: Deficiency of NLRP3 expression in NLRP3-/- mice contributes to the pathogenesis of chronic inflammation associated with cystitis through IRAKM.

Metal-Organic Framework Composite Photothermal Membrane for Removal of High-Concentration Volatile Organic Compounds from Water via Molecular Sieving.

Water-soluble volatile organic compounds (VOCs) are among the most difficult-to-treat species during wastewater treatment. The current purification and removal of high-concentration VOCs still rely on the energy-consuming distillation and high-pressure driven reverse osmosis technology. There is an urgent need for an advanced technology that can effectively remove high-concentration VOCs from water. Here, we report a metal-organic framework (MOF)/polyaniline (PANI) nanofiber array composite photothermal membrane for removal of high-concentration VOCs from water via molecular sieving during a solar-driven evaporation process. The modified zeolitic imidazole framework-8 (ZIF-8) layer grown on a PANI nanofiber array acts as a molecular sieving layer to evaporate water but intercept VOCs. The composite membrane exhibits high VOCs rejection and a high-water evaporation rate for water containing different concentrations of VOCs. When treating water containing VOCs with a concentration of up to 400 mg L-1, the VOCs rejection rate is up to 99% and the water evaporation rate is 1.0 kg m-2 h-1 under 1 sun irradiation (1 kW m-2). Our work effectively combines the molecular sieve effect with a solar-driven evaporation process, which provides an effective strategy for the treatment of water containing VOCs.

Normal prostate-derived stromal cells stimulate prostate cancer development.

Stromal cells play a decisive role in regulating tumor progression. In this study, we assessed the significance of normal prostate-derived stromal cells (PSCs) in prostate cancer development. An in vivo s.c. tumor model was established as follows: Group 1, DU145 cells alone; Group 2, DU145 + PSCs; Group 3, DU145 cells alone injected into pre-castrated mice; and Group 4, DU145 + PSCs injected into pre-castrated mice. Following injection, tumors were only detectable in the first two groups, with more aggressive growth in Group 2 than in Group 1 (P < 0.05). Immunohistochemical analysis revealed significantly higher proliferation (P < 0.05), but not apoptosis or altered expression of androgen receptor in Group 2, as compared with Group 1. In vitro, DU145 cells isolated from Group 1 tumors showed lower viability and migratory capability than those from Group 2. cDNA microarray on isolated DU145 cells from Groups 1 and 2 revealed the differential expression of genes regulating cell cycle progression and cell mobility, including GADD45A, RHOV, KLK11, and PCK1. Our results suggest that stromal cells derived from normal prostate potentiate the development of tumor growth in vivo, which is achieved at least in part through the regulation of cell-cycle- and migration-related gene expression within the tumor cells.

Establishment of a Risk Score Model for Early Prediction of Severe H1N1 Influenza.

H1N1 is the most common subtype of influenza virus circulating worldwide and can cause severe disease in some populations. Early prediction and intervention for patients who develop severe influenza will greatly reduce their mortality. In this study, we conducted a comprehensive analysis of 180 PBMC samples from three published datasets from the GEO DataSets. Differentially expressed gene (DEG) analysis and weighted correlation network analysis (WGCNA) were performed to provide candidate DEGs for model building. Functional enrichment and CIBERSORT analyses were also performed to evaluate the differences in composition and function of PBMCs between patients with severe and mild disease. Finally, a risk score model was built using lasso regression analysis, with six genes (CX3CR1, KLRD1, MMP8, PRTN3, RETN and SCD) involved. The model performed moderately in the early identification of patients that develop severe H1N1 disease.

Bipolar transurethral resection in saline vs traditional monopolar resection of the prostate: results of a randomized trial with a 2-year follow-up.

OBJECTIVE: To present 2-year follow-up data of a randomized clinical trial comparing bipolar transurethral resection in saline (TURIS) with monopolar transurethral resection of the prostate (TURP). PATIENTS AND METHODS: In all, 100 consecutive patients with benign prostatic hyperplasia (BPH) were randomized to TURIS or TURP. The breath ethanol test was used before and every 10 min during surgery to assess fluid absorption. Complications and treatment efficacy were evaluated after surgery. RESULTS: The operative duration and resected tissue weight were similar between the groups. The mean decreases in serum sodium and haemoglobin after surgery were significantly less in the TURIS group. The mean (standard deviation) irrigant absorbed was significantly less in TURIS than in the TURP group, at 208 (344) mL vs 512 (706) mL respectively (P < 0.001). In both the TURIS and TURP groups there were significant improvements in International Prostate Symptoms Scores and maximum urinary flow rates. The acute and late complications in the groups were statistically similar. CONCLUSION: Bipolar TURIS seems to be a safe and effective procedure, which is associated with significantly less fluid absorption and similar efficacy as traditional monopolar TURP.

[Expressions of ATF3 and CTGF and their regulation by estradiol in the prepuce of hypospadias patients].

OBJECTIVE: Estrogen is closely associated with hypospadias. The present study was to explore the molecular mechanism of hypospadias caused by estradiol. METHODS: Fibroblasts obtained from the prepuce of hypospadiac and normal children were cultured in vitro and treated with 17-beta ethinyl estradiol (17-EE) at the concentrations of 1 micromol/L to 0.1 nmol/L for 2 hours, or at 0.1 micromol/L for 0.5, 1, 2, 4, 8, 16 and 24 hours. MTT assay was used to evaluate the effect of 17-EE on the proliferation of the cells, and RT-PCR was employed to detect the expressions of the activating transcription factor-3 (ATF3) and connective tissue growth factor (CTGF) in the hypospadiac tissue. The results were compared with those obtained from the nonhypospadiac tissue. RESULTS: The expressions of ATF3 and CTGF were significantly upregulated in the hypospadiac tissue as compared with the nonhypospadiac group. At the concentration of 1 micromol/L, 17-EE significantly inhibited the proliferation of the cells. ATF3 mRNA was elevated at 1-2 hours, while CTGF mRNA showed no significant changes in 24 hours. CONCLUSION: ATF3 and CTGF are two candidate genes involved in the etiology of hypospadias. And estradiol may induce hypospadias by upregulating the expressions of ATF3 and CTGF.

Multifunctional hollow superhydrophobic SiO2 microspheres with robust and self-cleaning and separation of oil/water emulsions properties.

Superhydrophobic materials have drawn great attention due to its' remarkable non-wetting properties and applications in many fields. In this paper, we synthesize a hollow superhydrophobic SiO2 powder by typical template method and self-assembly functionalization. Robustness of many superhydrophobic surfaces has become the development bottleneck for industrial applications. Aiming at this problem, the adhesive epoxy resin is specially taken to use as the binding layer between superhydrophobic SiO2 powder and substrates to create robust superhydrophobic coating. The mechanical durability of the obtained superhydrophobic coating is evaluated by a cyclic sandpaper abrasion. Also, the chemical stability of this superhydrophobic coating is assessed by exposuring it to different pH conditions and UV irradiation, respectively. Significantly, because of the special structure and superhydrophobicity/superoleophilicity of the hollow microspheres, these hollow superhydrophobic SiO2 powders manifest great oil-adsorbing capacity, which thus can be used to separate oil/water mixtures and remove oil from oil-in-water emulsions.

Adiponectin deficiency contributes to the development and progression of benign prostatic hyperplasia in obesity.

The incidence of benign prostatic hyperplasia (BPH) is increasing among obese individuals, but few studies have fully explained the underlying mechanisms. We aimed to elucidate the relationship between obesity and BPH. Herein, we show that in prostatic epithelial and stromal cells, adiponectin exerts multifunctional effects including anti-proliferation, blocking of G1/S-phase progression and the promotion of apoptosis via inhibiting the MEK-ERK-p90RSK axis. Furthermore, we found that a high-fat diet (HFD) led to adiponectin deficiency and microscopic BPH in a mouse model of obesity. And an adiponectin supplement protected the obese mice from microscopic BPH. The present study provides evidence that adiponectin is a protective regulator in the development and progression of BPH and that adiponectin deficiency causally links BPH with obesity.

The effects of RNA interference mediated VEGF gene silencing on biological behavior of renal cell carcinoma and transplanted renal tumor in nude mice.

OBJECTIVE: This study was to explore the effects of RNA interference mediated vascular endothelial growth factor (VEGF) gene silencing on biological behavior of renal cell carcinoma (RCC), transplanted renal tumor and angiogenesis in nude mice. METHODS: The specific siRNA sequence targeting VEGF were designed and synthesized to construct hVEGF-siRNA plasmid which was transfected into RCC 786-O cells. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used for the detection of VEGF gene expression and western blot was adopted for the examination of VEGF protein expression. The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect cell growth as well as cell migration and invasion. The transplanted renal tumor models in nude mice were established, and the growth condition of nude mice, and VEGF protein expression in transplanted tumor slices and the microvessel density (MVD) were detected. RESULTS: The expression level of VEGF mRNA in VEGF-siRNA group was significant lower than that in the control group and negative group, suggesting that establishment of plasmid specifically inhibited the expression of VEGF gene The expression level of VEGF protein in VEGF-siRNA group was significant lower than that in the control group and negative group. VEGF gene silencing has the significant inhibition effects on proliferation, migration and invasion of RCC 786-O cells. The tumor weight, VEGF protein positive rate and MVD in VEGF-siRNA group were significant lower than those in negative group and blank group. CONCLUSION: The VEGF gene silencing could inhibit the cell proliferation, migration and invasion of RCC 786-O cells; inhibition of VEGF protein expression could prevent transplanted RCC growth and tumor angiogenesis.

Differential effects of peripheral and transitional prostatic stromal cells on tumorigenesis.

The human prostate contains two types of stromal cells, peripheral stromal cells (PSCs) and transitional stromal cells (TSCs). Here, we demonstrate the effects of PSCs and TSCs on tumorigenesis in prostate cancer (PCa) and identify the mechanisms underlying these effects. Using microarray analysis, we identified 3,643 differentially expressed genes in cocultures of TSCs, PSCs, and DU145 cells, a human prostate cancer cell line. Expression of cell division cycle 25 homolog A (CDC25A) was lower and that of tumor-associated calcium signal transducer 2 (TACSTD2) was higher in TSCs than in PSCs. Additionally, increased CDC25A expression or decreased TACSTD2 expression modulated the survival, growth, and migration of DU145 cells. These data suggest that PSCs promote and TSCs inhibit tumorigenesis by regulating the expression of CDC25A and TACSTD2.

Human Stromal Cells in the Peripheral Zone of the Prostate Promote Tumorigenesis of Prostatic Cancer Stem Cells through Up-regulation of C-Kit Expression.

OBJECTIVE: Most prostate cancers originate from the prostatic peripheral zone (PZ). We tested the hypothesis that the stromal cells from PZ and transitional zone (TZ) have differential effects on the ability of tumorigenesis. METHODS: Stromal cells isolated from the PZ and TZ of normal human prostates mixed with DU145 cells subcutaneously injected into athymic nude mice. The volume and weight of tumors was measured and analyzing the ability of purified DU145 cells isolated from the tumors to migrate and proliferate. The expression patterns of stem cell-specific genes of these DU145 cells were examined. The C-Kit inhibitor, imatinib mesylate, was administrated to confirm the effect of stromal cells on the tumorigenesis. RESULTS: The volume and weight of tumors were significantly higher in mice transplanted with DU145 and stromal cells from PZ. In contrast, the data was significantly lower with DU145 and stromal cells from TZ than DU145 alone. The purified DU145 cells isolated from the tumors with DU145 and stromal cells in PZ had increased ability to migrate and proliferate, and had increased expression of C-Kit. These effects of the stromal cells in the PZ on DU145 cells could be blocked using imatinib mesylate. CONCLUSIONS: Human stromal cells in the PZ promote the in vivo tumorigenesis of DU145 through up-regulating C-Kit; in contrast, the stromal cells in the TZ inhibit it through down-regulating the expression of C-Kit. The model will be useful for understanding the mechanisms by which the prostatic stem cell niche controls the tumorigeneis of prostatic cancer stem cells.

Normal peripheral prostate stromal cells stimulate prostate cancer development: roles of c-kit signal.

BACKGROUND: To investigated the peripheral stromal cell conditioned medium (CM) -stimulated c-kit-JAK2-STAT1 pathway in prostate cancer. METHODS: CM harvested from normal prostate peripheral stromal cells was added to DU145 cells. DU145 cell viability and migration were measured by cell counting kit-8 reagent and Transwell analysis respectively. Colony and sphere formation efficiencies of DU145 cells co-cultured with CM from human prostate stromal cells were also measured. DU145cells were stably transfected with lentivirus-mediated shRNA for c-kit silencing. RESULTS: C-kit expression in prostate cancer was found to be significantly higher than in benign prostatic hyperplasia and positively associated with Gleason scores. The growth, migration and capacity of clonogenic property of DU145 cells significantly increased upon exposure to peripheral stromal CM and then were inhibited after silencing the expression of c-kit. The levels of c-kit, pJAK2 and pSTAT1 were significantly induced by peripheral zone stromal CM compared with controls in serum free medium and the levels of pJAK2 and pSTAT1 decreased after c-kit silencing. CONCLUSIONS: C-kit hyper-expression promotes the development of prostate cancer. The peripheral stromal cell CM stimulated c-kit-JAK2-STAT1 pathway in prostate cancer cell viability, migration, and capacity of clonogenic property. This may lead to a greater understanding of the role of c-kit in prostate cancer and provide a potential therapeutic target for prostate cancer.

RNAi-mediated knockdown of vascular endothelial growth factor inhibits vascularization and tumor growth in renal cell carcinoma.

In the present study, we aimed to examine the effects of the knockdown of vascular endothelial growth factor (VEGF) by RNA interference (RNAi) on vascularization and tumor growth in renal cell carcinoma (RCC). For this purpose, a lentiviral vector expressing VEGF-shRNA was constructed and transfected into 293T cells. The efficiency of RNAi was determined by infecting human 786-O RCC cells with viral particles and measuring the VEGF mRNA levels by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The effect of transfection with VEGF­shRNA on the secreted VEGF levels was also examined and the inhibitory effects on vascularization were also examined using a chick chorioallantoic membrane (CAM) assay. An RCC xenograft model was established in nude mice by implanting 786-O cells to form subcutaneous tumors. VEGF expression was observed by immunohistochemical (IHC) staining of the xenograft tumors. The tumor volume and tumor inhibition rate were also recorded. The apoptosis of the cancer cells was measured by TUNEL assay and the efficiency of tumor inhibition was estimated. The interference rate of VEGF­shRNA was 72.2% in the 786-O cells. Our results revealed that VEGF mRNA expression, the secreted VEGF level in the 786-O cells and the total vessel length were markedly reduced in the VEGF­shRNA-transfected cells compared with the controls (all P<0.05). Compared with the controls, injections of lentivirus expressing VEGF-shRNA significantly inhibited tumor growth, and reduced tumor mass and VEGF expression in the tumor tissue (all P<0.05). The apoptotic index in the treatment group was significantly higher than that in the controls (both P<0.05). Thus, our data indicate that the inhibition of VEGF expression by RNAi reduces VEGF mRNA levels, and inhibits angiogenesis and tumor growth in RCC, providing a future treatment option for RCC.

Increased Expression of GOLPH3 is Associated with the Proliferation of Prostate Cancer.

BACKGROUND: Golgi phosphoprotein 3 (GOLPH3) is a metastasis-associated gene, however its role in cell proliferation of prostate cancer (PCa) has not yet been elucidated. METHODS: The level of expression of GOLPH3 and other genes was examined by quantitative real-time PCR (QPCR) and western blot analysis. Furthermore, we performed a comprehensive analysis of the expression of GOLPH3 in PCa using a tissue microarray (TMA) and correlated our findings with pathological parameters of PCa. RNA interference (RNAi) was used to silence the expression of GOLPH3 in PC-3 cells and to measure the effects on proliferation and cell cycle using the CCK-8 assay and flow cytometry. Western blots were also employed to assess AKT-mTOR and cell cycle-related proteins. RESULTS: We showed that the expression of GOLPH3 was located at the trans-Golgi membranes in PCa cells. We found that GOLPH3 was expressed in all PCa cells and was significantly higher in two androgen-independent cell lines, DU145 and PC-3. TMA immunohistochemistry showed that GOLPH3 was positive in 64% of cancer tissue samples compared with 20% in normal and 30% in benign samples (P<0.05). In vitro, silencing GOLPH3 expression inhibited cell proliferation and arrested the cell cycle at the G2/M phase. Silencing GOLPH3 also activated P21 expression but suppressed the expression of CDK1/2 and cyclinB1 protein together with the phosphorylation of AKT and mTOR. CONCLUSIONS: The expression of the GOLPH3 protein was significantly elevated in PCa. GOLPH3 can promote cell proliferation by enhancing the activity of AKT-mTOR signaling. Altogether, these findings suggest that GOLPH3 play important roles in proliferation and cell cycle regulation in PCa and might serve as promising biomarkers for PCa progression as well as potential therapeutic targets.