Cancer-Associated Fibroblasts Regulate Bladder Cancer Invasion and Metabolic Phenotypes through Autophagy.

Recently, both cancer-associated fibroblasts (CAFs) and autophagy have been proven to play an important role in tumor development, including bladder cancer (BCa). However, the real mechanisms remain largely unclear. Here, we reconstruct a mimic tumor microenvironment to explore the interaction between CAFs and the BCa cell line T24 using a coculture system. Autophagy in CAFs was induced or inhibited by rapamycin or siRNA, respectively. After coculture with CAFs, T24 cell proliferation, invasion, and aerobic glycolysis were tested in vitro. Rapamycin induced and siAtg5 inhibited autophagy in CAFs. Enhanced autophagy in CAFs promoted cell proliferation and invasion in T24 cells in vitro, while there was no significant difference between the autophagy-inhibited group and the controls. Lactate concentration was elevated in both rapamycin-treated and siAtg5-treated groups compared with the control group. In addition, the expression levels of MCT1, MCT4, HK2, SLC2A1, and MMP-9 were all increased in T24 cells in the autophagy-enhanced group. Our results indicated that CAFs could regulate BCa invasion and metabolic phenotypes through autophagy, providing us with new alternative treatments for BCa in the future.

Identification and verification of an immune-related lncRNA signature for predicting the prognosis of patients with bladder cancer.

BACKGROUND: Recent studies have revealed the significant roles of immune-related long noncoding RNAs (lncRNAs) in cancer development and progression. The identification of biomarkers that contribute to early detection and risk stratification provides significant benefits for bladder cancer (BC) patients. The current study aimed to determine an immune-related lncRNA signature for predicting the prognosis of BC patients. METHODS: Based on The Cancer Genome Atlas (TCGA) database, we identified seven immune-related lncRNAs with prognostic value. The predictive value of the prognostic signature developed from immune-related lncRNAs was assessed by survival and nomogram analyses. Principal component analysis (PCA) was performed to visualize gene expression patterns in the groups defined by the risk score, and the immune composition and purity of the tumor were evaluated by the ESTIMATE algorithm. RESULTS: Based on the Pearson correlation analysis results, 765 immune-related lncRNAs were filtered (|R| > 0.4, P < 0.001), and seven immune-related lncRNAs (Z84484.1, AC009120.2, AL450384.2, AC024060.1, TNFRSF14-AS1, AL354919.2, OCIAD1-AS1) with prognostic value were finally identified. Patients in the low-risk group had a better prognosis than those in the high-risk group. Multivariate Cox regression analysis showed that the signature was an independent prognostic factor. A prognostic nomogram with clinical features and the signature of seven immune-related lncRNAs was also constructed. According to the PCA and ESTIMATE algorithm results, we found different immune statuses in the low-and high-risk groups. CONCLUSIONS: Our study shows that the signature of seven immune-related lncRNAs can be used as a prognostic marker for BC.

Elucidation of the Mechanism by Which a ADAMTS5 Gene MicroRNA-Binding Site Single Nucleotide Polymorphism Affects the Risk of Osteoarthritis.

Purpose: The aim of this study was to investigate the relationship between single nucleotide polymorphisms (SNPs) within the 3' untranslated region of the microRNA (miRNA)-binding site of the ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5) gene and the risk of knee osteoarthritis (KOA) and its mechanism. Materials and Methods: Sanger sequencing was used to determine the genotypes of three ADAMTS5 gene SNPs (rs3171407, rs229071, and rs229077) from 310 patients suffering from osteoarthritis of the knee joint (KOA) and 310 healthy controls. The levels of the miRNAs, thsa-miR-103b, hsa-miR-144-3p, and hsa-miR-105-5p in plasma, and the level of ADAMTS5 mRNA in the articular cartilage of 132 patients with KOA were detected by real-time quantitative PCR. Results: The G allele at the rs3171407 locus of the ADAMTS5 gene was demonstrated to be a protective genetic factor for KOA. In addition, the risk of developing KOA was significantly increased in subjects carrying the C allele at the rs229071 locus. The risk of developing KOA in carriers of the G allele at locus rs229077 was 1.49 times greater than those with the A allele. The level of the hsa-miR-144-3p was increased, and the expression level of the ADAMTS5 protein was decreased in carriers of the T allele at the rs229071 locus. In carriers of rs229077 locus A allele the hsa-miR-105-5p level was increased and the expression level of ADAMTS5 protein was decreased. Conclusion: SNPs at the rs3171407, rs229071, and rs229077 loci of the ADAMTS5 gene are related to the risk of OA. The likely mechanism underlying these observations is that these SNPs affect the regulation of ADAMTS5 protein expression through miRNAs; however, this needs to be verified using in vivo models.

Development and validation of a prognostic immune-associated gene signature in clear cell renal cell carcinoma.

BACKGROUND: Recent studies have demonstrated that immune-associated genes (IAGs) play an important role in the occurrence and progression of clear renal clear cell carcinoma (ccRCC). Novel biomarkers and a reliable prognostic prediction model for ccRCC patients are still limited. The objective of this study was to develop a IAGs signature and validate its prognostic value in ccRCC using bioinformatic methods and publicly database. METHODS: In the present study, we identified differentially expressed IAGs in ccRCC based on The Cancer Genome Atlas (TCGA) database. A prognostic IAGs risk model was further developed and its prognostic and predictive value was evaluated by survival analysis and nomogram. RESULTS: A total of 681 differentially expressed IAGs were identified and seven IAGs (IFI30, WNT5A, IRF9, AGER, PLAUR, TEK, BID) were finally selected in a IAGs signature. Survival analysis revealed that high IAGs risk scores were significantly related to poor survival outcomes. The IAGs signature was demonstrated as an independent prognostic factor and closely related to the metastasis status of ccRCC. A nomogram with clinicopathologic characteristics and IAGs signature was also constructed to superiorly predict prognosis of ccRCC patients. CONCLUSIONS: We identified seven IAGs as a potential signature for reflecting the prognosis of ccRCC based on TCGA database. Further clinical trials are needed to validate our observations and the mechanisms underlying the prognostic value of IAGs signature in ccRCC also deserve further experimental exploration.

Prognostic Value of Lymphovascular Invasion in Node-Negative Upper Urinary Tract Urothelial Carcinoma Patients Undergoing Radical Nephroureterectomy.

PURPOSE: This study aimed to evaluate the prognostic impact of lymphovascular invasion (LVI) in patients treated with radical nephroureterectomy (RNU) for upper urinary tract urothelial carcinoma (UTUC). MATERIALS AND METHODS: We collected data from 180 patients who were treated with RNU from 2005 to 2013 at our institution. The Kaplan-Meier method with log-rank test and Cox proportional hazards regression models were used for univariate and multivariate analyses. RESULTS: LVI was present in 28 patients (15.6%), which was associated with higher pathological tumor stage (p<0.001), tumor necrosis (p=0.012), lymph node metastasis (p=0.017) and multifocality (p=0.012). On multivariate analysis, LVI was an independent prognostic factor of recurrence-free survival [RFS: hazard ratio (HR)=2.954; 95% confidence interval (CI)=1.539-5.671; p=0.001] and cancer-specific survival (CSS: HR=3.530; 95% CI=1.701-7.325; p=0.001) in all patients. In patients with node-negative UTUC, LVI was also a significant predictor of RFS (HR=3.732; 95% CI 1.866-7.464; p<0.001) and CSS (HR=3.825; 95% CI=1.777-8.234; p=0.001). CONCLUSION: LVI status was an independent predictor in patients with UTUC who underwent RNU. The estimate of LVI could help physicians identify high-risk patients and make a better medication regimen of adjuvant chemotherapy.

MCT1 regulates aggressive and metabolic phenotypes in bladder cancer.

Background: Monocarboxylate transporter isoform 1 (MCT1) is an important molecule in mediating lactate transportation. Recent studies have shown an oncogenic role of MCT1 in cancer development. Methods: In this study, we aimed to investigate the expression and role of MCT1 in bladder cancer (BCa). MCT1 expression was detected in 124 BCa tissues and their clinicopathological significance was analyzed. We also used The Cancer Genome Atlas database to explore the prognostic association of MCT1 with BCa. Cell proliferation, migration and invasion assays were performed on BCa cells in which MCT1 was downregulated. The effect of MCT1 on BCa cell aerobic glycolysis, as well as its association with HIF-1α, was tested. Results: We found that high MCT1 expression correlated with lymph node and distant metastasis. Patients with high-MCT1 expression showed shorter overall survival than those with low-MCT1 expression. Knockdown of MCT1 inhibited BCa cell proliferation, migration and invasion, and affected expression of epithelial-mesenchymal transition related proteins. Downregulation of MCT1 decreased lactate levels in cell medium, as well as HK2, GLUT1 and LDHB expression. In addition, MCT1 expression was partly dependent on HIF-1α. Conclusions: Taken together, our study has shown a prognostic role of MCT1 in BCa, and provided potential diagnostic and therapeutic options for BCa patients.

Clusterin promotes growth and invasion of clear cell renal carcinoma cell by upregulation of S100A4 expression.

BACKGROUND AND OBJECTIVE: Clusterin promotes cell proliferation, motility and invasiveness in human renal cell carcinoma (RCC) cells but the underlying molecular mechanisms of this action are largely unknown. The aim of this study was to investigate the effects of clusterin on cancer cell growth, invasion and S100A4 expression and to determine the effects of clusterin on in vitro cell proliferation and migration and in vivo tumour growth in RCC cells. METHODS: We have established stable transfectants of highly invasive Caki-1 human RCC cells with expression of clusterin shRNA targeting clusterin (Caki-1/clusterin shRNA). We also established stable transfectants of 786-O human RCC cells with expression of clusterin cDNA plaismid (786-O/clusterin cDNA). Clusterin and S100A4 expression was detected by reverse transcription (RT) PCR and western blot assay; Caki-1/clusterin shRNA and 786-O/clusterin cDNA clones were subjected to in vitro-invasion assays. Cell viability and cell growth was assessed in MTT and clonogenic assay. Specific small interfering RNA was employed to down-regulate S100A4. The expression plasmid for S100A4 (pCMV-S100A4) was used to upregulate S100A4. Caki-1/clusterin shRNA clones were injected subcutaneously in nude mice to determine tumour growth and cancer cell invasiveness in vivo. Xenograft tumour tissues were assessed by immunohistochemistry and frozen tissues were used for the detection of S100A4 and clusterin. RESULTS: Overexpression of clusterin increased cell invasiveness; and targeting clusterin reduced cell invasiveness in vitro. This increase in cell invasiveness was mediated by S100A4. Targeting clusterin decreased cell proliferation and down-regulated cellular S100A4 levels in Caki-1 cells; Overexpression of clusterin increased cell proliferation and up-regulated cellular S100A4 levels in 786-O cells; Stable Caki-1/clusterin shRNA transfectants produced smaller xenograft tumours containing reduced S100A4 protein levels in vivo. Stable 786-O/clusterin cDNA transfectants produced larger xenograft tumours containing increased S100A4 protein levels in vivo. CONCLUSION: Our results indicate that clusterin promotes growth and invasion in RCC cells in vitro and in vivo through upregulation of S100A4; And targeting clusterin confers growth inhibitory and anti-invasive properties in RCC cells in vitro and in vivo through a down-regulation of S100A4. These findings provide the rationale for future oncostatic strategies aimed at suppressing clusterin-mediated signal transduction pathways as a novel therapeutic approach in human RCC.

Sex Differences in the Association between Night Shift Work and the Risk of Cancers: A Meta-Analysis of 57 Articles.

OBJECTIVES: To identify the association between night shift work and the risk of various cancers with a comprehensive perspective and to explore sex differences in this association. METHODS: We searched PubMed, Embase, and Web of Science for studies on the effect of night shift work on cancer, including case-control, cohort, and nested case-control studies. We computed risk estimates with 95% confidence intervals (CIs) in a random or fixed effects model and quantified heterogeneity using the I 2 statistic. Subgroup, metaregression, and sensitivity analyses were performed to explore potential sources of heterogeneity. Contour-enhanced funnel plots and the trim and fill method were used together to analyze bias. Linear dose-response analysis was used to quantitatively estimate the accumulative effect of night shift work on the risk of cancer. RESULTS: Fifty-eight studies were eligible for our meta-analysis, including 5,143,838 participants. In the random effects model, the pooled odds ratio (OR) of cancers was 1.15 (95% CI = 1.08-1.22, P < 0.001; I 2 = 76.2%). Night shift work increased the cancer risk in both men (OR = 1.14, 95% CI = 1.05-1.25, P = 0.003) and women (OR = 1.12, 95% CI = 1.04-1.20, P = 0.002). Subgroup analyses showed that night shift work positively increased the risk of breast (OR = 1.22, 95% CI = 1.08-1.38), prostate (OR = 1.26, 95% CI = 1.05-1.52), and digestive system (OR = 1.15, 95% CI = 1.01-1.32) cancers. For every 5 years of night shift work, the cancer risk increased by 3.2% (OR = 1.032, 95% CI = 1.013-1.051). CONCLUSION: This is the first meta-analysis identifying the positive association between night shift work and the risk of cancer and verifying that there is no sex difference in the effect of night shift work on cancer risk. Cancer risk increases with cumulative years of night shift work.

Protective effects of the AKT activator SC79 on renal ischemia-reperfusion injury.

BACKGROUND AND AIMS: SC79, an AKT activator, has been reported to protect experimental ischemia-elicited neuronal death, brain injury, and myocardiocyte hypoxia/reoxygenation (H/R) injury. However, the protection of SC79 from renal ischemia-reperfusion (I/R) injury and the precise mechanisms involved are unknown. Here, we investigated the effects of SC79 in renal tubular epithelial cells in vitro and in mouse kidney in vivo following hypoxia-reoxygenation (H/R) and renal I/R injury. METHODS: The kidneys of Sprague-Dawley rats were subjected to 30 min of warm ischemia followed by 24 h of reperfusion. Murine renal tubular epithelial NRK-52E cells were subjected to hypoxia for 6 h and reoxygenation for 24 h. The NRK-52E cells and the renal I/R injury model were treated with SC79 and/or LY294002 at different times and concentrations. Serum creatinine (Cr) concentration, renal histology, cellular viability, and cell apoptosis were assessed. Levels of phospho-Akt, bad, Bim, bax, bcl-2, and bcl-XL in NRK-52E cells and renal tissues were determined by western blotting. RESULTS: SC79 improved viability and inhibited apoptosis in NRK-52E cells following H/R. SC79 decreased serum Cr and markedly improved pathology and decreased cell apoptosis in kidneys following I/R. In addition, SC79 promoted the expression of phospho-Akt, bcl-2, and bcl-XL, and decreased the expression levels of bid, bax, and bim. PI3K inhibitor (LY294002) pre-treatment completely abolished these effects of SC79. CONCLUSIONS: The protective role of SC79 against H/R of NRK-52E cells or renal I/R injury is related to activation of phosphorylation of AKT, resulting in a decrease in the pro-apoptotic proteins bim, bax, and bad and an increase in the anti-apoptotic proteins bcl-2 and bcl-XL induced by cell H/R and renal I/R injury.

MicroRNA-126 inhibits tumor cell invasion and metastasis by downregulating ROCK1 in renal cell carcinoma.

MicroRNAs (miRNAs) are involved in cancer development and progression. Renal cell carcinoma (RCC) frequently undergoes metastasis and has a high mortality rate. The current study measured miRNA­126 (miR­126) expression levels in 128 pairs of clear cell RCC and adjacent normal kidney tissue samples by reverse transcription­quantitative polymerase chain reaction, and analyzed the association between miR­126 and various clinicopathological parameters. In addition, cell proliferation, wound healing and cell invasion assays were conducted using RCC cells overexpressing miR­126. Potential miR­126 target genes and the signaling pathways that may be regulated by miR­126 were then examined. miR­126 expression was significantly reduced in patients with metastatic RCC compared with patients without metastasis. Consistently, overexpression of miR­126 in RCC cells significantly inhibited cell proliferation, migration and invasion in vitro compared with negative control miRNA. A luciferase reporter assay demonstrated that miR­126 targets Rho associated coiled­coil containing protein kinase 1 (ROCK1) by directly binding the 3'­untranslated region. Furthermore, western blotting identified miR­126 as an important regulator of the AKT and extracellular signal­regulated 1/2 signaling pathways. The results of the present study indicate that miR­126 inhibits RCC cell proliferation, migration and invasion by downregulating ROCK1. These findings suggest that miR­126 may be valuable as a potential target for therapeutic intervention in RCC.

The association between metabolic syndrome and the risk of urothelial carcinoma of the bladder: a case-control study in China.

BACKGROUND: Evidence of the association of metabolic syndrome (MetS) with cancer risk is accumulating. However, uncertainties still exist as to the link of MetS with bladder cancer. This study aimed to assess the relationship between MetS and the risk of urothelial carcinoma of the bladder (UC) in a Chinese population. METHODS: We retrospectively analyzed clinicopathological data of 972 newly diagnosed UC patients and 1098 cancer-free controls matched to the cases by age and gender. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using unconditional logistic regression in both unadjusted and adjusted models. RESULTS: MetS was not significantly associated with the overall UC risk (p=0.08). However, a significant association of MetS with UC was observed in female patients (p=0.006). Diabetes mellitus (crude OR 1.339, 95% CI 1.079-1.662, p=0.008; adjusted OR 1.767, 95% CI 1.308-2.386, p<0.001) and hypertriglyceridemia (crude OR 1.245, 95% CI 1.018-1.522, p=0.033; adjusted OR 1.254, 95% CI 1.020-1.542, p=0.032) were significantly associated with UC risk. As the number of MetS components increased, the UC risk was elevated. Having three or more (versus zero) components of MetS was significantly related to risk of overall UC (OR 1.315; 95% CI 1.006-1.719; p=0.045) and non-muscle invasive bladder cancer (OR 1.354; 95% CI 1.019-1.798; p=0.037). CONCLUSIONS: The present study indicated a marginal association between MetS and UC risk, and a significant association with UC risk in female patients. The results need to be evaluated in large-scale prospective cohorts.

ADIPOQ polymorphism rs182052 is associated with clear cell renal cell carcinoma.

Recent studies have indicated that low circulating adiponectin concentrations are associated with a higher risk of several cancers, including renal cell carcinoma. In this case-control study, we examined the frequency of single nucleotide polymorphisms (rs182052G>A, rs266729C>G, and rs3774262G>A) in the adiponectin gene (ADIPOQ) in 1004 patients with clear cell renal cell carcinoma (ccRCC) compared with a group of healthy subjects (n = 1108). Fasting serum adiponectin concentrations were also examined. Logistic regression models were used to estimate odds ratios (OR) and 95% confidence intervals (95% CI). The association of serum adiponectin concentration with genetic variants was calculated using a multivariate linear regression model. A significantly higher ccRCC risk was associated with the rs182052 variant A allele (adjusted OR, 1.36 and 95% CI, 1.07-1.74 for AA vs GG, P = 0.013; adjusted OR, 1.27 and 95% CI, 1.04-1.56 for AA vs GG+AG, P = 0.019), and this positive association was more evident in overweight subjects. Fasting serum adiponectin was lower in subjects carrying A alleles of rs182052 in both ccRCC patients (ß = -0.399, P = 0.018) and healthy controls (ß = -0.371, P = 0.024). These results suggest that ADIPOQ rs182052 is significantly associated with ccRCC risk.

The association between metabolic syndrome and advanced prostate cancer in Chinese patients receiving radical prostatectomy.

The global incidence of metabolic syndrome (MetS) is dramatically increasing. Considerable interest has been devoted to the relationship between MetS and prostate cancer (PCa) risk. However, few studies have examined the association between MetS and PCa progression. This retrospective study consisted of 1016 patients with PCa who received radical prostatectomy. The association between MetS and pathological features was evaluated using logistic regression analysis. Compared with patients without MetS, those with MetS indicated an increased risk of prostatectomy Gleason score (GS) ≥8 (odds ratio [OR] =1.670, 95% confidence interval (CI) 1.096-2.545, P= 0.017), and a 1.5-fold increased risk of pT3-4 disease (OR = 1.583, 95% CI 1.106-2.266, P= 0.012). The presence of MetS was an independent predictor of lymph node involvement (OR = 1.751, 95% CI 1.038-2.955, P= 0.036). Furthermore, as the number of MetS components accumulated, the risk of a GS ≥ 8 increased. The present study indicates a significant association between MetS and advanced PCa. The results need to be evaluated in large-scale prospective cohorts.

Clinical outcome of advanced and metastatic renal cell carcinoma treated with targeted therapy: is there a difference between young and old patients?

BACKGROUND: To assess whether the clinical outcome of advanced and metastatic renal cell carcinoma (mRCC) treated with targeted therapy differs between young and old patients. PATIENTS AND METHODS: A total of 327 patients with advanced renal cell carcinoma and mRCC who received targeted therapy in two Chinese clinical centers were analyzed retrospectively. The patients were stratified into three groups: young (aged <45 years), middle-aged (aged 45-64 years), and old (aged ≥65 years). Overall survival (OS) and progression-free survival (PFS) curves were drawn using the Kaplan-Meier method, and Cox's proportional hazard regression model was used to compare OS and PFS within age groups. RESULTS: There were no significant differences among young, middle-aged, and old groups in terms of OS (P=0.087), whereas PFS in the old group was significantly better than in the young and middle-aged groups (P=0.043). Both OS and PFS in the younger groups (aged <65 years) were significantly worse than in the old group (age ≥65 years; median OS, 28.1 vs 28.7 months [P=0.029]; median PFS, 11.4 vs 14 months [P=0.015]). No difference in OS or PFS was found between the young and middle-aged groups. After adjusting for sex, body mass index, smoking status, hypertension, diabetes mellitus, Eastern Cooperative Oncology Group score, history of cytokines, and Fuhrman grade, old age was an independent favorable prognostic factor for OS and PFS compared with younger age (<65 years) (OS, hazard ratio, 0.552 [95% confidence interval, 0.329-0.828; P=0.006]; PFS, hazard ratio, 0.584 [95% confidence interval, 0.401-0.850; P=0.005]). CONCLUSION: Younger patients with advanced renal cell carcinoma and mRCC receiving targeted therapy have a poorer prognosis compared with old patients. These results remain to be examined in prospective cohorts.

Prognostic significance of the epithelial-to-mesenchymal transition markers e-cadherin, vimentin and twist in bladder cancer.

OBJECTIVE: The goal of this study was to utilize long-term patient follow-up to determine whether epithelial-to-mesenchymal transition (EMT)-related markers can predict bladder cancer patient survival and progression of disease. MATERIALS AND METHODS: This study included 121 patients with bladder cancer. Sixty-four of these patients presented with non-muscle invasive (NMI, stage T1) bladder cancer and 57 with muscle invasive (MI, stage T2, T3). The patients were diagnosed and treated between May 1998 and July 2012. The EMT markers E-cadherin, Twist, and Vimentin were detected via immunohistochemistry. Univariate and multivariate/Cox analyses were then utilized to determine whether these EMT markers could be useful prognostic markers for predicting bladder cancer patient outcomes. RESULTS: Analysis of the 121 bladder cancer patients in this study revealed that the frequency of E-cadherin expression was 59.5% (72/121), Twist was 54.5% (66/121), and Vimentin was 24.8% (30/121). Twist and Vimentin were found to have statistically significant correlations with grade, recurrence, and progression but not with stage, whereas E-cadherin was associated with stage but not with the other parameters. In the univariate analysis, grade (p = 0.02) was the only significant predictor for progression-free survival (PFS). Stage, grade, and expression of E-cadherin, Vimentin and Twist were included in the multivariate analysis of predicting PFS. In this analysis, grade (p = 0.01) and Vimentin expression (p = 0.001) were found to be significant prognostic factors in predicting PFS. CONCLUSIONS: Grade and Vimentin are potential independent indicators in predicting bladder cancer progression and survival.

Prognostic significance of the epithelial-to-mesenchymal transition markers e-cadherin, vimentin and twist in bladder cancer

ObjectiveThe goal of this study was to utilize long-term patient follow-up to determine whether epithelial-to-mesenchymal transition (EMT)-related markers can predict bladder cancer patient survival and progression of disease.Materials and MethodsThis study included 121 patients with bladder cancer. Sixty-four of these patients presented with non-muscle invasive (NMI, stage T1) bladder cancer and 57 with muscle invasive (MI, stage T2, T3). The patients were diagnosed and treated between May 1998 and July 2012. The EMT markers E-cadherin, Twist, and Vimentin were detected via immunohistochemistry. Univariate and multivariate/Cox analyses were then utilized to determine whether these EMT markers could be useful prognostic markers for predicting bladder cancer patient outcomes.ResultsAnalysis of the 121 bladder cancer patients in this study revealed that the frequency of E-cadherin expression was 59.5% (72/121), Twist was 54.5% (66/121), and Vimentin was 24.8% (30/121). Twist and Vimentin were found to have statistically significant correlations with grade, recurrence, and progression but not with stage, whereas E-cadherin was associated with stage but not with the other parameters. In the univariate analysis, grade (p = 0.02) was the only significant predictor for progression-free survival (PFS). Stage, grade, and expression of E-cadherin, Vimentin and Twist were included in the multivariate analysis of predicting PFS. In this analysis, grade (p = 0.01) and Vimentin expression (p = 0.001) were found to be significant prognostic factors in predicting PFS.ConclusionsGrade and Vimentin are potential independent indicators in predicting bladder cancer progression and survival.

Clusterin plays an important role in clear renal cell cancer metastasis.

OBJECTIVE: Clusterin (CLU) is implicated in regulating clear renal cell carcinoma (CRCC) progression and metastasis, yet the mechanisms are not elucidated. In the present study, we explored the potential role of CLU in CRCC metastasis. METHODS: Levels of CLU mRNA and CLU protein were measured by RT-PCR and immunohistochemistry analysis in 22 CRCC with metastasis and 22 without metastasis and 22 samples of normal kidney tissue. After CLU silencing and re-expression, the migration and invasion in vitro and in vivo of Caki-2 cells were determined by wound healing assay, transwell migration assay and pulmonary nodule assay, respectively. The expression of pERK1/2 and MMP-9 were detected by RT-PCR and Western blot assay. RESULTS: We found a significant increase of CLU and CLU mRNA expression in CRCC, and the expression of CLU is strongly correlated in patients with metastatic disease. We discovered that CLU-rich Caki-2 cells displayed higher invasive ability which prompted us to investigate if CLU silencing could reduce the migration and invasion in Caki-2 cells. Compared with the vector-transfected cells, CLU knocked-down (CLUi) cells showed reduced migration and invasion in vitro, as well as decreased metastatic potential in experimental metastasis. Re-expression of CLU in CLUi cells restored the invasive phenotypes. We found that MMP-9 was downregulated in CLUi cells. We also discovered that levels of activated ERK1/2 correlated with the rich expression of CLU and MMP-9. CONCLUSION: Our data suggest that CLU may regulate aggressive behavior of human CRCC cells through modulating ERK1/2 signaling and MMP-9 expression.

Cytochrome P450 2C8 and flavin-containing monooxygenases are involved in the metabolism of tazarotenic acid in humans.

Upon oral administration, tazarotene is rapidly converted to tazarotenic acid by esterases. The main circulating agent, tazarotenic acid is subsequently oxidized to the inactive sulfoxide metabolite. Therefore, alterations in the metabolic clearance of tazarotenic acid may have significant effects on its systemic exposure. The objective of this study was to identify the human liver microsomal enzymes responsible for the in vitro metabolism of tazarotenic acid. Tazarotenic acid was incubated with 1 mg/ml pooled human liver microsomes, in 100 mM potassium phosphate buffer (pH 7.4), at 37 degrees C, over a period of 30 min. The microsomal enzymes that may be involved in tazarotenic acid metabolism were identified through incubation with microsomes containing cDNA-expressed human microsomal isozymes. Chemical inhibition studies were then conducted to confirm the identity of the enzymes potentially involved in tazarotenic acid metabolism. Reversed-phase high performance liquid chromatography was used to quantify the sulfoxide metabolite, the major metabolite of tazarotenic acid. Upon incubation of tazarotenic acid with microsomes expressing CYP2C8, flavin-containing monooxygenase 1 (FMO1), or FMO3, marked formation of the sulfoxide metabolite was observed. The involvement of these isozymes in tazarotenic acid metabolism was further confirmed by inhibition of metabolite formation in pooled human liver microsomes by specific inhibitors of CYP2C8 or FMO. In conclusion, the in vitro metabolism of tazarotenic acid to its sulfoxide metabolite in human liver microsomes is mediated by CYP2C8 and FMO.