YAP1 inhibits RSL3-induced castration-resistant prostate cancer cell ferroptosis by driving glutamine uptake and metabolism to GSH.

High levels of YAP1 and ferroptosis activation in castration-resistant prostate cancer (CRPC) can inhibit CRPC progression and improve its sensitivity toward chemotherapeutics drugs. However, whether YAP1 regulates ferroptosis in CRPC cells and the underlying mechanisms are unknown. The protein levels of YAP1, SLC1A5, and GLS1 in benign prostatic hyperplasia (BPH), prostate cancer (PCa) that did not progress to CRPC, and CRPC tissue samples were evaluated using western blotting. In PC-3 and DU-145 cells, YAP1 overexpression vector, small-interfering RNA, specific inhibitor verteporfin, ferroptosis-inducer RSL3, SLC1A5-inhibitor V-9302, and GLS1-inhibitor CB-839 were used. Immunofluorescence, flow cytometry, dual-luciferase reporter gene, and related kits were used to investigate the effect of YAP1 on the ferroptosis activity in CRPC cells and its underlying mechanisms. YAP1 promoted extracellular glutamine uptake and subsequent production of glutamate and glutathione (GSH), and increases the GPX4 activity. For the activation of ferroptosis by RSL3, YAP1 decreased the levels of reactive oxygen species, malondialdehyde, and lipid peroxidation, and the proportion of dead cells. Mechanistically, YAP1 promoted the expression of SCL1A5 and GLS1 and further increased the GSH levels and GPX4 activity. Thus, inhibiting SLC1A5 or GLS1 activity could alleviate the antagonistic effect of YAP1 on the ferroptosis of RSL3-induced CRPC cells. In CRPC, the YAP1 level is high, which enters the nucleus and promotes the expressions of SLC1A5 and GLS1, thereby promoting cellular glutamine uptake and metabolism to generate glutamate and further synthesizing GSH, increasing GPX4 activity, improving cellular antioxidant capacity, and inhibiting cell death.

HO-1 Protects Remnant Liver against Dysfunction after Major Hepatectomy in Humans.

OBJECTIVE: During major resection of liver carcinoma, liver regeneration (LR) is induced by various clinical and biological factors. Heme oxygenase (HO)-1 has been found as a key inducer of LR in preclinical trial. The clinical evidence for the role of HO-1 in liver dysfunction (LD) including LR is still unknown and has been included in this study. METHODS: Therefore, plasma HO-1 were monitored during perioperative period in 65 patients with hepatectomy, with 35 training and 30 validation cohorts. LD were evaluated by liver function serum markers and calculation of regeneration indices, respectively. RESULTS: In the training setting, HO-1 levels were remarkably reduced after liver resection (P < 0.001) and gradually recovered within 7 days after surgery. Preoperative HO-1 specifically predicted LD during the first week after surgery (AUC: 0.757; P = 0.01). In patients with LD and complications after surgery, HO-1 levels decreased throughout the perioperative period. In addition, we had also confirmed that low levels of HO-1 (<169 ng/ml) before surgery were associated with an increase in the incidence of postoperative LD and morbidity (P = 0.007, P = 0.045), and decrease of liver regeneration (P = 0.005). And HO-1 was an independent predictor for poor clinical outcome. CONCLUSIONS: We had provided the first clinical data verifying that human HO-1 was related to LD. Consequently, HO-1 levels can be used as effective clinical indicators to predict LD and clinical outcome, and can be used as intervention target before liver resection.

Circular RNA CircPPP1CB Suppresses Tumorigenesis by Interacting With the MiR-1307-3p/SMG1 Axis in Human Bladder Cancer.

Circular RNA (circRNA) is a newly discovered endogenous non-coding RNA (ncRNA), which is characterized with a closed circular structure. A growing body of evidence has verified the vital roles of circRNAs in human cancer. In this research, we selected circPPP1CB as a study object by circRNA sequencing and quantitative real-time PCR (qRT-PCR) validation in human bladder cancer (BC). CircPPP1CB is downregulated in BC and is negatively correlated with clinical stages and histological grades. Functionally, circPPP1CB modulated cell growth, metastasis, and epithelial-to-mesenchymal transition (EMT) process in vitro and in vivo. Mechanically, we performed various experiments to verify the circPPP1CB/miR-1307-3p/SMG1 regulatory axis. Taken together, our results demonstrated that circPPP1CB participates in tumor growth, metastasis, and EMT process by interacting with the miR-1307-3p/SMG1 axis, and that circPPP1CB might be a novel therapeutic target and diagnostic biomarker in human BC.

Thymoquinone Suppresses the Proliferation, Migration and Invasiveness through Regulating ROS, Autophagic Flux and miR-877-5p in Human Bladder Carcinoma Cells.

Bladder carcinoma is among the top 10 most frequently diagnosed cancer types in the world. As a phytochemical active metabolic, thymoquinone (TQ) is extracted from seeds of Nigella sativa, possessing various biological properties in a wide range of diseases. Moreover, the outstanding anti-cancer effect of TQ is attracting increasing attentions. In certain circumstances, moderate autophagy is regarded to facilitate the adaptation of malignant cells to different stressors. Conversely, closely linked with the mitochondrial membrane potential (MMP) loss, the upregulation of intracellular reactive oxygen species (ROS) is reported to activate the cell apoptosis in many cancer types. Furthermore, the vital effects of microRNAs in the pathological processes of cancer cells have also been confirmed by previous studies. The present research confirms that TQ restrains the viability, proliferation, migration and invasion through activating caspase-dependent apoptosis in bladder carcinoma cells, which is mediated by TQ induced ROS increase in bladder carcinoma cells. Furthermore, TQ is proved to block the fusion of autophagosomes and lysosomes, causing the accumulation of autophagosomes and subsequent cell apoptosis. In addition, TQ is also found to initiate the miR-877-5p/PD-L1 axis, which suppresses the epithelial mesenchymal transition (EMT) and invasion of bladder carcinoma cells. Taken together, TQ induces the apoptosis through upregulating ROS level and impairing autophagic flux, and inhibiting the EMT and cell invasion via activating the miR-877-5p/PD-L1 axis in bladder carcinoma cells.

A positive feedback loop between TAZ and miR-942-3p modulates proliferation, angiogenesis, epithelial-mesenchymal transition process, glycometabolism and ROS homeostasis in human bladder cancer.

BACKGROUND: Transcriptional coactivator with PDZ-binding motif (TAZ) has been reported to be involved in tumor progression, angiogenesis, epithelial-mesenchymal transition (EMT), glycometabolic modulation and reactive oxygen species (ROS) buildup. Herein, the underlying molecular mechanisms of the TAZ-induced biological effects in bladder cancer were discovered. METHODS: qRT-PCR, western blotting and immunohistochemistry were performed to determine the levels of TAZ in bladder cancer cells and tissues. CCK-8, colony formation, tube formation, wound healing and Transwell assays and flow cytometry were used to evaluate the biological functions of TAZ, miR-942-3p and growth arrest-specific 1 (GAS1). QRT-PCR and western blotting were used to determine the expression levels of related genes. Chromatin immunoprecipitation and a dual-luciferase reporter assay were performed to confirm the interaction between TAZ and miR-942. In vivo tumorigenesis and colorimetric glycolytic assays were also conducted. RESULTS: We confirmed the upregulation and vital roles of TAZ in bladder cancer. TAZ-induced upregulation of miR-942-3p expression amplified upstream signaling by inhibiting the expression of large tumor suppressor 2 (LATS2, a TAZ inhibitor). MiR-942-3p attenuated the impacts on cell proliferation, angiogenesis, EMT, glycolysis and ROS levels induced by TAZ knockdown. Furthermore, miR-942-3p restrained the expression of GAS1 to modulate biological behaviors. CONCLUSION: Our study identified a novel positive feedback loop between TAZ and miR-942-3p that regulates biological functions in bladder cancer cells via GAS1 expression and illustrated that TAZ, miR-942-3p and GAS1 might be potential therapeutic targets for bladder cancer treatment.

Circular RNA circRIMS1 Acts as a Sponge of miR-433-3p to Promote Bladder Cancer Progression by Regulating CCAR1 Expression.

Circular RNAs (circRNAs), a subclass of noncoding RNAs, are reportedly involved in the progression of various diseases. However, the exact role of circRIMS1, also termed hsa_circ_0132246, in human bladder cancer remains unknown. By performing RNA sequencing comparing bladder cell lines and normal uroepithelial cells, circRIMS1 was selected as a research object. We further verified by qRT-PCR that circRIMS1 is upregulated in both bladder cancer tissue and cell lines. Proliferation, colony-formation, Transwell migration, invasion, apoptosis, western blotting, and in vivo experiments were utilized to clarify the roles of circRIMS1, microRNA (miR)-433-3p, and cell cycle and apoptosis regulator 1 (CCAR1). For mechanistic investigation, RNA pulldown, fluorescence in situ hybridization (FISH), and luciferase reporter assay confirmed the binding of circRIMS1 with miR-433-3p. Inhibition of circRIMS1 suppressed the proliferation, migration, and invasion of bladder cancer cells both in vitro and in vivo. Moreover, the circRIMS1/miR-433-3p/CCAR1 regulatory axis was confirmed to be responsible for the biological functions of circRIMS1. Taken together, our research demonstrated that circRIMS1 promotes tumor growth, migration, and invasion through the miR-433-3p/CCAR1 regulatory axis, representing a potential therapeutic target and biomarker in bladder cancer.

Artesunate induces autophagy dependent apoptosis through upregulating ROS and activating AMPK-mTOR-ULK1 axis in human bladder cancer cells.

Artesunate is a kind of derivative of artemisinin, which possesses potent anti-cancer effect in addition to its anti-malarial property. And autophagy was a highly conserved process, exerting a double-edged effect in cancer cell survival. Besides, apoptosis is a programmed cell death program, crucial to cell homeostasis. However, the relations between autophagy and apoptosis, and the role of artesunate in this interaction have not been elucidated in bladder cancer. In present study, we used human bladder cancer cells (T24 and EJ cell lines) to investigate that how artesunate would influence autophagy and apoptosis processes. We found that artesunate could inhibit the viability, proliferation and migration of bladder cancer cells, as well as induce autophagy in a time and dose dependent manner, in addition, the artesunate induced autophagy subsequently activated cells apoptosis. Furthermore, we pretreated T24 and EJ cells with 3-Methyladenine or Rapamycin to inhibit or promote autophagy, respectively, leading to inhibited or increased apoptosis. Moreover, pretreatment of these cell lines with Acadesine or Dorsomorphin to activate or inhibit the AMPK-mTOR-ULK1 pathway, respectively, also resulting in promotion or suppression in both autophagy and apoptosis. In the upstream, ROS upregulation triggered by ART initiated AMPK-mTOR-ULK1 axis. However, this initiative effect of ROS can be reversed by N-Acetyl-l-cysteine. Therefore, this study indicated that Artesunate induces autophagy dependent apoptosis through upregulating ROS and activating AMPK-mTOR-ULK1 pathway in human bladder cancer cells.

Ureteroscopic Diagnosis and Povidone Iodine Treatment for Chronic Unilateral Hematuria Caused by Benign Lesions.

BACKGROUND The management of chronic unilateral hematuria (CUH) caused by benign lesions is a therapeutic challenge to many urologists. The aims of this study were to evaluate the efficacy and safety of povidone iodine sclerotherapy for CUH. MATERIAL AND METHODS We identified 20 patients who underwent povidone iodine sclerotherapy to treat CUH between September 2013 and August 2017. Radiologic and hematologic tests were normal, no definite cause of hematuria was revealed, and the malignant lesions were excluded. Cystoscopy and ureteroscopy indicated the lesions were located in the renal pelvis. The goal of successful treatment was no recurrence of hematuria during follow-up. RESULTS The present study analyzed 20 patients (9 females and 11 males), 24-73 years old (mean age 44.6) with mean follow-up of 23.8 (range 13-60) months. Endoscopic findings included discrete lesions, diffuse lesions, and no obvious lesion. Discrete lesions were identified as hemangioma (4/20, 20%), minute venous rupture (12/20, 60%), and varix (1/20, 5%). Diffuse lesions were founded via ureteroscopy in 2 (2/20, 10%) patients. In the remaining 1 (1/20, 5%) patient, no obvious lesion was found. All patients with CUH were treated with 0.5% povidone iodine for pelvicalyceal system instillation, which was given at 12-h intervals for 3 days. Only 1 patient experienced recurrent gross hematuria, after 24 months postoperatively. The overall success rate, defined as resolution of gross hematuria after povidone iodine sclerotherapy, was 95%. No complications were recorded. CONCLUSIONS The present study indicates that povidone iodine sclerotherapy could be an effective, safe, and minimally invasive treatment for chronic unilateral hematuria caused by benign lesions.

Sodium butyrate inhibits migration and induces AMPK-mTOR pathway-dependent autophagy and ROS-mediated apoptosis via the miR-139-5p/Bmi-1 axis in human bladder cancer cells.

Bladder cancer is one of the most frequently occurring malignant tumors in the urinary system. Sodium butyrate (NaB) is a histone deacetylase inhibitor and exerts remarkable antitumor effects in various cancer cells. MicroRNAs (miRNAs) and autophagy play crucial roles in cancer occurrence and development. In the present study, we evaluated the anticancer effects, including cell migration inhibition and the apoptotic effects of NaB in human bladder cancer cells. Furthermore, we found that NaB inhibited migration and induced AMPK/mTOR pathway-activated autophagy and reactive oxygen species (ROS) overproduction via the miR-139-5p/Bmi-1 axis. In addition, we found that ROS overproduction contributed to NaB-induced caspase-dependent apoptosis and autophagy. The interplay between autophagy and apoptosis in NaB treatment was clarified. Our findings provide a further understanding of EMT reversion, apoptosis and autophagy induced by antitumor drugs and a novel perspective and alternative strategy for bladder cancer chemotherapy.

AMPK-mTOR-ULK1 axis activation-dependent autophagy promotes hydroxycamptothecin-induced apoptosis in human bladder cancer cells.

10-hydroxycamptothecin (HCPT), a natural plant extract, exerts anticancer capacity. HCPT has been reported to induce apoptosis and autophagy in human cancer cells. The interaction between autophagy and apoptosis induced by HCPT and the molecular mechanism in bladder cancer cells were investigated in this study. Our results confirmed that HCPT suppressed cell viability and migration and caused cell-cycle arrest in T24 and 5637. Then, we used Z-VAD(OMe)-FMK to clarify that apoptosis induced by HCPT was mediated by caspase. Moreover, HCPT boosted autophagy through activating the AMPK/mTOR/ULK1 pathway. Blocking autophagy by 3-methyladenine, the adenosine monophosphate-activated protein kinase (AMPK) inhibitor dorsomorphin and siATG7 reversed HCPT-induced cytotoxicity. Conversely, rapamycin and the AMPK activator AICAR enhanced growth inhibition and cell apoptosis, suggesting that autophagy played a proapoptosis role. Taken together, our findings showed that HCPT-induced autophagy mediated by the AMPK pathway in T24 and 5637 cell lines, which reinforced the apoptosis, indicating that HCPT together with autophagy activator would be a novel strategy for clinical treatment in bladder cancer.

[Microsurgical subinguinal varicocelectomy with delivery of the testis and ligation of gubernacular veins: Evaluation of clinical effects].

OBJECTIVE: To compare the clinical effects and postoperative complications of microsurgical subinguinal varicocelectomy (MSV) with or without delivery of the testis and ligation of gubernacular veins in the treatment of varicocele. METHODS: We retrospectively analyzed the clinical data about 163 varicocele patients treated by MSV, 40 with (group A) and the other 123 without delivery of the testis and ligation of gubernacular veins (group B). We compared the operation time, postoperative complications, rate of recurrence, and semen parameters before and at 3 months after surgery between the two groups of patients. RESULTS: The operation time was significantly longer in group A than in B (ï¼»81.1 ± 20.0ï¼½ vs ï¼»62.3 ± 9.6ï¼½ min, P = 0.041). Sperm concentration, total sperm count per ejaculate, sperm viability, and the percentage of progressively motile sperm were significantly improved in both groups at 3 months after MSV as compared with the baseline (P < 0.05). There were no statistically significant differences in the above semen parameters between the two groups of patients with grade Ⅲ varicocele before and after surgery (P < 0.05). Scrotal edema developed in 5 cases in group A and wound infection in 2 cases in group B after MSV, but no postoperative testicular atrophy or recurrence was observed in either of the two groups. CONCLUSIONS: MSV with delivery of the testis and ligation of gubernacular veins showed no advantages over that without in reducing varicocele recurrence and improving semen parameters, but rather involved longer operation time and a higher incidence rate of postoperative complications.

Prostatic Calculi: Do They Matter?

INTRODUCTION: Prostatic calculi (PC) are frequently detected at computed tomography or ultrasound in men attending the health center or the urology outpatient department. PC have attracted more attention from urologists, but the clinical significance of PC is unknown. AIM: To review the available literature on the effects of PC on prostatic diseases and sexual function in men. METHODS: Relevant clinical trials were identified by searching the PubMed, Embase, and Cochrane Library databases. Results were classified, summarized, and analyzed. MAIN OUTCOME MEASURES: Transabdominal and rectal ultrasonography; urodynamics analysis; International Prostate Symptom Score; pathologic examination of prostatic tissue; prostate-specific antigen; and expressed prostatic secretion. RESULTS: PC can not only prolong the duration of bothersome symptoms but also decrease the cure rate of antibacterial therapy in patients with chronic prostatitis. Patients with PC usually have more severe lower urinary tract symptoms (LUTS), and some studies reported that moderate to marked PC are a predisposing factor for moderate to severe LUTS. Studies also reported that the serum level prostate-specific antigen is not influenced by PC. In addition, the presence of PC is not associated with an increased risk of prostate cancer. However, the correlation between PC in the peripheral zone and prostate cancer is statistically significant. In addition, the association between PC and Gleason scores is controversial. Some novel studies suggested that PC might play an important role in sexual impairment in middle-age men or men with chronic pelvic pain syndrome or chronic prostatitis. Recently, PC were found to increase the incidence of severe LUTS, urinary retention, and hematospermia after transrectal ultrasound-guided prostate biopsy. CONCLUSION: PC can aggravate LUTS, chronic prostatitis, and sexual dysfunction in men, but the association between PC and prostate cancer is still controversial. Cao J-J, Huang W, Wu H-S, et al. Prostatic Calculi: Do They Matter? Sex Med Rev 2018;6:482-491.

Risk factors for bladder calculi in patients with benign prostatic hyperplasia.

We aim to find the risk factors that influence the formation of bladder calculi in patients with benign prostate hyperplasia (BPH) and to reduce the surgical intervention related to bladder calculi.Between January 2015 and October 2016, 332 patients with BPH underwent surgical therapy were retrospectively evaluated. Patients with BPH were categorized into 2 groups: 94 patients with bladder calculi in group 1 and 238 patients without bladder calculi in group 2. Medical history, age, body mass index (BMI), total prostate specific antigen, total prostate volume (TPV), International Prostate Symptom Score (IPSS), intravesical prostatic protrusion (IPP), urodynamic parameters, and urine culture were compared between groups.There was no significant difference in the age, BMI, peak flow rate, and total IPSS between groups. TPV, total prostate specific antigen, and duration of BPH were significantly lower in group 1 than those in group 2. In addition, IPP was significantly higher in group 1 than group 2 (P < .001). Besides, after exclusion of patients with urinary retention and indwelling catheter, group 1 associated with a significantly higher preoperative positive rate of urine culture than that of group 2 (P = .046). Multivariate analysis indicated that IPP was a significant independent risk factor for the presence of bladder calculi.The incidence of bladder calculi in patients with BPH was proved to be closely associated with preoperative positive urine culture and longer IPP in our study. Furthermore, the IPP was presented to be an independent risk factor for the formation of bladder calculi. And early antibacterial therapy of urinary tract infection (UTI) may help to prevent the presence of bladder calculi in patients with BPH.