Novel Reversible-Binding PET Ligands for Imaging Monoacylglycerol Lipase Based on the Piperazinyl Azetidine Scaffold.

Monoacylglycerol lipase (MAGL) is a 33 kDa serine protease primarily responsible for hydrolyzing 2-arachidonoylglycerol into the proinflammatory eicosanoid precursor arachidonic acid in the central nervous system. Inhibition of MAGL constitutes an attractive therapeutic concept for treating psychiatric disorders and neurodegenerative diseases. Herein, we present the design and synthesis of multiple reversible MAGL inhibitor candidates based on a piperazinyl azetidine scaffold. Compounds 10 and 15 were identified as the best-performing reversible MAGL inhibitors by pharmacological evaluations, thus channeling their radiolabeling with fluorine-18 in high radiochemical yields and favorable molar activity. Furthermore, evaluation of [18F]10 and [18F]15 ([18F]MAGL-2102) by autoradiography and positron emission tomography (PET) imaging in rodents and nonhuman primates demonstrated favorable brain uptakes, heterogeneous radioactivity distribution, good specific binding, and adequate brain kinetics, and [18F]15 demonstrated a better performance. In conclusion, [18F]15 was found to be a suitable PET radioligand for the visualization of MAGL, harboring potential for the successful translation into humans.

Design, Synthesis, and Evaluation of Reversible and Irreversible Monoacylglycerol Lipase Positron Emission Tomography (PET) Tracers Using a "Tail Switching" Strategy on a Piperazinyl Azetidine Skeleton.

Monoacylglycerol lipase (MAGL) is a serine hydrolase that degrades 2-arachidonoylglycerol (2-AG) in the endocannabinoid system (eCB). Selective inhibition of MAGL has emerged as a potential therapeutic approach for the treatment of diverse pathological conditions, including chronic pain, inflammation, cancer, and neurodegeneration. Herein, we disclose a novel array of reversible and irreversible MAGL inhibitors by means of "tail switching" on a piperazinyl azetidine scaffold. We developed a lead irreversible-binding MAGL inhibitor 8 and reversible-binding compounds 17 and 37, which are amenable for radiolabeling with 11C or 18F. [11C]8 ([11C]MAGL-2-11) exhibited high brain uptake and excellent binding specificity in the brain toward MAGL. Reversible radioligands [11C]17 ([11C]PAD) and [18F]37 ([18F]MAGL-4-11) also demonstrated excellent in vivo binding specificity toward MAGL in peripheral organs. This work may pave the way for the development of MAGL-targeted positron emission tomography tracers with tunability in reversible and irreversible binding mechanisms.

EORTC risk tables are more suitable for Chinese patients with nonmuscle-invasive bladder cancer than AUA risk stratification.

BACKGROUND: Patients with non-muscle-invasive bladder cancer (NMIBC) need accurate estimations of the risk of recurrence and progression. Physicians can offer individualized therapy after identifying high-risk tumors. In our study, we compared the applicability of European Organization for Research and Treatment of Cancer (EORTC) risk tables and American Urological Association (AUA) risk stratification in Chinese patients with NMIBC. METHODS: We retrospectively studied 301 patients with NMIBC who underwent transurethral resection of bladder tumor (TURBT) between October 2000 and July 2009 at Huashan Hospital of Fudan University and analyzed their parameters. The recurrence and progression rates at 1 and 5 years postoperatively were calculated along with 95% confidence intervals. We compared them with results obtained from the EORTC risk tables and AUA risk stratification. P values <.05 were considered statistically significant. RESULTS: The median patient age was 67 years (21-92 years) and the median follow-up duration was 46 months (2-151 months). We used EORTC risk tables to classify patients into 3 groups, depending on whether they suffered recurrence or progression after TURBT. Kaplan-Meier curves showed significant differences among the 3 recurrence-free survival (RFS) levels (P < .0001, log-rank test) and among the 3 progression-free survival (PFS) levels (P < .0001, log-rank test). AUA risk stratification showed the same results. Both classifications were suitable to predict recurrence and progression in Chinese patients. However, for high-risk patients in both series, Kaplan-Meier curves showed significant differences between RFS levels (P < .0001, log-rank test) and between PFS levels (P < .0001, log-rank test). EORTC risk tables were stricter and AUA was more sensitive in assigning patients to a high-risk group. CONCLUSION: EORTC risk tables are better than AUA risk stratification for predicting recurrence and progression in Chinese patients with NMIBC, especially among high-risk patients.

LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway.

Prostate cancer (PCa) is the most commonly diagnosed malignancy in male. Numerous studies have focused on the molecular mechanisms of carcinogenesis and progression, aiming at developing new therapeutic strategies. Here we describe Lanthionine synthase C-like protein 1 (LanCL1), a member of the LanCL family, is a potential prostate cancer susceptibility gene. LanCL1 promotes prostate cancer cell proliferation and helps protect cells from damage caused by oxidative stress. Suppression of LanCL1 by siRNA results in increased cancer cell apoptosis. Clinical data also indicate that LanCL1 upregulation in human prostate cancers correlates with tumor progression. Finally, we demonstrate that LanCL1 plays such important role through inhibiting JNK pathway. Altogether, our results suggest that LanCL1 protects cells from oxidative stress, and promotes cell proliferation. LanCL1 reduces cell death via suppression of JNK signaling pathway.

Overexpression of WDFY2 inhibits prostate cancer cell growth and migration via inactivation of Akt pathway.

Prostate cancer is the most commonly diagnosed malignancy and is the second leading deadly reason among male cancer. WDFY2, which is found to be a cancer-specific fusion gene with CDKN2D in ovarian cancer, is a new gene with unknown function in carcinogenesis. In this study, we investigated the role of WDFY2 in prostate cancer development. We examined WDFY2 expression in human prostate tissue specimens and prostate cancer cell lines BPH-1, LNCaP, PC3, and DU-145. Overexpression of WDFY2 was performed to evaluate the role of WDFY2 in cell proliferation, migration, and colony formation of prostate cancer cells. We analyzed the clinical impact and prognosis of WDFY2 expression on the progress of prostate cancer through data from online datasets. Our results showed that WDFY2 had lower expression level in prostate tumors than in normal tissues. Overexpression of WDFY2 in prostate cancer cells DU145 and PC-3 led to the suppression of cancer cell migration and colony formation. Furthermore, we found that WDFY2 exerted its role by suppressing the activity of Akt pathway other than the epithelial-mesenchymal transition progression. In conclusion, we have uncovered WDFY2 as a tumor suppressor gene and a new potential biomarker for cancer progression. Our results showed that WDFY2 inhibited cancer cell colony formation and migration via suppressing Akt pathway, making it a potential new therapeutic target in prostate cancer.

Germline genetic variations in PDZD2 and ITPR2 genes are associated with clear cell renal cell carcinoma in Chinese population.

Genome-wide association studies (GWAS) of renal cell carcinoma (RCC) have identified single nucleotide polymorphisms (SNPs) associated with RCC in European and African American population. In this study, we evaluated whether these SNPs are associated with clear cell RCC (ccRCC) in Chinese population. All reported RCC risk-associated SNPs from GWAS were evaluated in 346 ccRCC cases and 1,130 controls. Rs10054504 (at PDZD2, Odds ratio, OR = 0.71, 95%CI:0.59-0.86, P = 0.0006), rs718314 (at ITPR2, OR = 0.56, 95%CI:0.45-0.69, P = 5.26×10-8) and rs1049380 (at ITPR2, by dominant model, OR = 1.58, 95%CI:1.18-2.13, P = 0.0025) were significantly associated with ccRCC risk in Chinese population. To conclude, genetic variations in PDZD2 and ITPR2 are ccRCC-risk associated in Chinese population.

Genetic scores based on risk-associated single nucleotide polymorphisms (SNPs) can reveal inherited risk of renal cell carcinoma.

The objective of this study was to evaluate whether renal cell carcinoma (RCC) risk-associated single nucleotide polymorphisms (SNPs) could reflect the individual inherited risks of RCC. A total of 346 RCC patients and 1,130 controls were recruited in this case-control study. Genetic scores were calculated for each individual based on the odds ratios and frequencies of risk-associated SNPs. Four SNPs were significantly associated with RCC in Chinese population. Two genetic score models were established, genetic score 1 (rs10054504, rs7023329 and rs718314) and genetic score 2 (rs10054504, rs7023329 and rs1049380). For genetic score 1, the individual likelihood of RCC with low (<0.8), medium (0.8-1.2) and high (≥1.2) genetic score 1 was 15.61%, 22.25% and 33.92% respectively (P-trend=6.88×10(-7)). For genetic score 2, individual with low (<0.8), medium (0.8-1.2) and high (≥1.2) genetic score 2 would have likelihood of RCC as 14.39%, 24.54% and 36.48%, respectively (P-trend=1.27×10(-10)). The area under the receiver operating curve (AUC) of genetic score 1 was 0.626, and AUC of genetic score 2 was 0.658. We concluded that genetic score can reveal personal risk and inherited risk of RCC, especially when family history is not available.

Periostin Mediates TGF-ß-Induced Epithelial Mesenchymal Transition in Prostate Cancer Cells.

BACKGROUND: In our previous study, we found that periostin was upregulated in prostate cancer, and its expression could be modulated by TGF-ß. TGF-ß could upregulate periostin expression in some cells, and both TGF-ß and periostin could induce epithelial mesenchymal transition (EMT). We aimed to study the effect of periostin in the process of TGF-ß-induced EMT in prostate cancer cells. METHODS: We constructed a lentivirus vector containing the periostin gene and transduced it into PC3 and DU145 cells. After confirming periostin overexpression by PCR and Western blotting, we used an MTT assay to establish a growth curve to measure cell proliferation. Additionally, we performed transwell and wound healing assays to measure cell invasion and migration, respectively. Lastly, we measured the expression of EMT associated factors using Western blot analysis to test the effect of periostin on EMT in prostate cancer cells. RESULTS: PCR and Western blot analyses confirmed that periostin was upregulated after infection with the periostin lentiviral vector. Periostin overexpression promoted increased cell proliferation, invasion, and migration as measured by MTT, transwell, and wound healing assays, respectively. Western blot analysis illustrated that periostin overexpression increased the expression of EMT associated factors, and periostin overexpression activated Akt and GSK-3ß, which could be inhibited using a PI3K inhibitor. Additionally, TGF-ß increased the levels of STAT3, Twist1 and periostin, while both STAT3 shRNA and Twist1 shRNA inhibited periostin expression. However, STAT3 shRNA also decreased Twist1 expression. Although reduction of STAT3, Twist1 or periostin levels with shRNA inhibited TGF-ß-induced overexpression of EMT associated factors, periostin overexpression could reverse such inhibition by interfering with STAT3 and Twist1. Similarly, periostin overexpression also reversed inhibition of cell invasion induced by interference of STAT3 and Twist1. CONCLUSION: Our findings indicate that periostin is an important mediator of TGF-ß-induced EMT and suggest that periostin is a potential therapeutic target for suppressing the metastatic progression of prostate cancer.

Human epidermal growth factor receptor 2: a significant indicator for predicting progression in non-muscle-invasive bladder cancer especially in high-risk groups.

PURPOSE: Current pathological and clinical parameters provide important prognostic information. However, they are still limitations for predicting the true malignant potential of a specific cancer. The aim of this study was to validate the predicting role of HER-2 expression and demonstrated that combination of the high-risk factors with HER-2 expression is more valuable for determining which non-muscle-invasive bladder cancer (NMIBC) is more aggressive. MATERIALS AND METHODS: In total, 238 patients treated by transurethral resection of the bladder tumor were histopathologically confirmed to be NMIBC. Two experienced uropathologists re-reviewed the slides. HER-2 expression was evaluated by immunohistochemistry and scored for intensity and area of staining. The association of HER-2 staining with tumor recurrence and progression was evaluated by univariate and multivariate analyses and Kaplan-Meier survival curves. RESULTS: In multivariable analyses, HER-2 expression was an independent risk factor for predicting tumor progression (HR 2.64, p = 0.024). Combining the EORTC risk scores with HER-2 expression status led to more accurate prediction of progression, especially in patients with intermediate- and high-risk EORTC scores (p < 0.0001, log-rank test). CONCLUSIONS: HER-2 positivity is prognostic for predicting progression to muscle invasion in NMIBC. Combination of the high-risk factors with HER-2 expression is more valuable for determining which NMIBC is more aggressive.

Snail is an independent prognostic indicator for predicting recurrence and progression in non-muscle-invasive bladder cancer.

OBJECTIVE: Snail, an inducer of the epithelial-to-mesenchymal transition, increases motility and invasiveness of cancer cells by repressing E-cadherin expression. We investigate the relationship between Snail expression and clinicopathological parameters and evaluate its prognostic significance in patients with non-muscle-invasive bladder cancer (NMIBC). MATERIALS AND METHODS: A total of 332 patients treated with transurethral resection of the bladder tumor between October 2002 and July 2010 were histopathologically confirmed to be NMIBC. Tumor recurrence and progression were followed up in all patients. Immunohistochemical staining of 332 slices was performed. The expression of Snail was evaluated by ICH and graded for intensity and area of staining. We correlated Snail scores with clinical and pathological variables, and association of Snail staining with tumor recurrence and progression was evaluated by univariate, multivariate analysis and Kaplan-Meier survival curves. RESULTS: Of 332 patients with NMIBC, there was Snail positivity in 104 tumors (31.3 %), and Snail expression correlated with age, multifocality, carcinoma in situ, tumor stage and tumor grade (each p < 0.05, respectively). A multivariate Cox regression model revealed that Snail expression was an independent predictor of tumor recurrence [hazard ratio (HR) 1.95, p = 0.001] and progression (HR 2.34, p = 0.014) in patients with NMIBC. Kaplan-Meier estimates showed that Snail expression was significantly associated with recurrence and progression (log-rank test, p < 0.0001, respectively). CONCLUSIONS: Analysis of Snail expression in 332 NMIBC tissue specimens revealed its potential usefulness as a biomarker to predict the NMIBC prognosis.

Retroperitoneal laparoscopic ureterolithotomy for proximal ureteral calculi in selected patients.

OBJECTIVES: To summarize our experience of retroperitoneal laparoscopic ureterolithotomy for ureteral calculi and evaluate the safety and efficiency of this procedure. METHODS: We conducted a retrospective analysis of 197 patients with proximal ureteral calculi who accepted retroperitoneal laparoscopic ureterolithotomy from June 2005 to June 2014. RESULTS: All procedures were performed successfully and the mean operating time and estimated blood loss were 87 min and 64 mL. The clearance rate was 98.5% and the rates of urine leak and ureteral stricture were 2.5% and 1.0%. CONCLUSIONS: Retroperitoneal laparoscopic ureterolithotomy is a safe and effective procedure for patients with complex stones or anatomic abnormalities, and, with experience of high volume series, it is also a reasonable choice as the primary treatment for such selected patients.

Are EORTC risk tables suitable for Chinese patients with non-muscle-invasive bladder cancer?

OBJECTIVE: To evaluate the applicability of using EORTC risk tables in Chinese patients with non-muscle-invasive bladder cancer. MATERIAL AND METHODS: Between October 2000 and July 2009, 301 patients with NMIBC who underwent transurethral resection of the bladder tumor (TURBT) at our hospital were followed up. The probability of recurrence and progression at 1 year and 5 years post-operatively was calculated along with the 95% confidence intervals. We then compared the actual probabilities in our center to those obtained through the application of the EORTC risk tables. RESULTS: Median patient age was 67 years (range, 21-92 years), and the median follow-up duration was 46 months (range, 2-151 months). The probability of recurrence at 1 year ranged from 2% to 58%, and the probability of progression ranged from less than 1.2% to 30%. At 5 years, the probability of recurrence ranged from 12% to 85%, and the probability of progression ranged from less than 2.9% to 50%. An overlapping of the confidence intervals of the probability between our series and the EORTC group is detected. CONCLUSIONS: Although the immediate instillation of intravesical chemotherapy may reduce the risk of recurrence, EORTC risk tables could predict recurrence and progression in Chinese patients with non-muscle-invasive bladder cancer.

Ki-67 is an independent indicator in non-muscle invasive bladder cancer (NMIBC); combination of EORTC risk scores and Ki-67 expression could improve the risk stratification of NMIBC.

OBJECTIVE: To prove the predicting role of Ki-67 expression and to demonstrate that the combination of European Organization for Research and Treatment of Cancer (EORTC) risk scores and Ki-67 staining status could improve the risk stratification in a large series of patients with non-muscle invasive bladder cancer (NMIBC). MATERIAL AND METHODS: From October 2002 to July 2010, in our cohort, 332 patients who were treated with transurethral resection of the bladder tumor were diagnosed with NMIBC by histopathologic analysis. Two experienced uropathologists rereviewed the slides. The EORTC risk scores for recurrence and progression were determined. Ki-67 expression was evaluated using immunohistochemical studies and scored for intensity and area of staining. We correlated Ki-67 expression scores with clinical and pathologic variables. We evaluated the prognosis role of EORTC risk scores, Ki-67 staining, and their combination on tumor recurrence-free survival and progression-free survival (PFS) by univariate analysis, multivariate analysis, and Kaplan-Meier survival curves. RESULTS: With a median follow-up of 47 (range, 2-124) months, 119 patients (35.8%) had tumor recurrence and 40 patients (12%) had tumor progression. Ki-67 positivity (Ki-67>25%) was reported in 108 tumors (32.5%), and it was significantly associated with high EORTC risk scores for both tumor recurrence and progression. In univariate analysis, multifocality, tumor size, tumor stage, tumor grade, and Ki-67 staining correlated with recurrence-free survival, whereas tumor size, tumor stage, tumor grade, concomitant CIS, and Ki-67 staining correlated with PFS. In multivariable analysis, Ki-67 expression was an independent risk factor for predicting tumor recurrence (hazard ratio, 2.14; P<0.0001) and progression (hazard ratio: 2.97, P = 0.004). Kaplan-Meier curves showed that combining EORTC risk scores and Ki-67 staining led to more accurate prediction for tumor recurrence and progression (log-rank test; P<0.0001). CONCLUSIONS: Ki-67 positivity is prognostic for predicting tumor recurrence and progression. Combination of EORTC risk scores with Ki-67 expression could improve the risk stratification for both recurrence and progression in NMIBC.

The prognostic value of C-reactive protein in renal cell carcinoma: a systematic review and meta-analysis.

OBJECTIVES: C-reactive protein (CRP) has been reported to be associated with poorer prognosis in patients with renal cell carcinoma (RCC); however, conflicting results exist. We conducted a systematic review to evaluate the prognostic value, and a meta-analysis was done if the extracted data could be merged. MATERIALS AND METHODS: We searched MEDLINE, EMBASE, and the Cochrane Central Search library for published studies that analyzed the effect of CRP in RCC. All included cases were categorized into 4 groups of different stages and tumor types for analysis, and the relationships between CRP and stage, grade, and survival were analyzed. RESULTS: Overall, 24 studies including 4,100 RCC cases were accepted for meta-analysis. Elevated CRP level was associated with higher stage (risk ratio [RR] 2.90, 95% confidence interval [CI] 2.52-3.32, P<0.00001) and higher grade (RR 4.31, 95% CI 3.35-5.56, P<0.00001) in the overall analysis of patients with all pathologic types of RCCs, and it was also associated with poorer overall survival (hazard ratio [HR] 1.51, 95% CI 1.09-1.93, P<0.00001) and cancer-specific survival (CSS) (HR 3.91, 95% CI 2.18-5.64, P<0.00001). In patients with localized RCC, elevated CRP level was associated with poorer CSS (HR 3.49, 95% CI 2.93-4.05, P<0.00001) and progression-free survival (HR 3.29, 95% CI 2.91-3.67, P<0.00001); whereas in patients with metastatic RCC, elevated CRP level was associated with poorer overall survival (HR 2.37, 95% CI 2.14-2.60, P<0.00001) and CSS (HR 3.70, 95% CI 3.19-4.22, P<0.00001). Specifically, in the patients with clear cell RCC, elevated CRP level was also associated with higher stage (RR 2.92, 95% CI 2.25-3.80, P<0.00001), poorer CSS (HR 2.60, 95% CI 2.32-2.88, P<0.00001), and poorer progression-free survival (HR 1.21, 95% CI 0.94-1.47, P<0.00001). CONCLUSION: Elevated CRP level in a patient with RCC is associated with poorer prognosis, and it could serve as a useful biomarker for clinical prediction.

Synthesis and characterization of superparamagnetic CoFe2O4/MWCNT hybrids for tumor-targeted therapy.

Owing to their great potentialities of carbon nanotubes (CNTs)-based magnetic nano-composites, numerous applications of them have been found in nanotechnology, integrated functional system, and in medicine. Herein, nearly monodisperse CoFe2O4 nanoparticles have been deposited on multi-walled carbon nanotubes (MWCNTs) by high-temperature hydrolysis and inorganic polymerization of ionic Co(II) and Fe(III) salts and MWCNTs in a polyol solution. X-ray diffraction, energy-dispersive X-ray spectrometry and transmission electron microscopy were used to characterize the final products. The average size of CoFe2O4 nanoparticles and their coverage density on MWCNTs can be adjusted to some extent by altering the reaction parameters. A proposed formation mechanism of the magnetic hybrids is presented. Magnetic measurements showed that the hybrids were superparamagnetic at room temperature and their saturation magnetization could be fine tuned by changing the loading of CoFe2O4 nanoparticles on the MWCNTs.

A systematic review and meta-analysis of current evidence comparing laparoendoscopic single-site adrenalectomy and conventional laparoscopic adrenalectomy.

PURPOSE: To assess the efficacy and safety of laparoendoscopic single-site adrenalectomy (LESS-A) and conventional laparoscopic adrenalectomy (LA) as a systematic review and meta-analysis of current evidence. METHODS: We conducted a thorough search for comparative studies that compared LESS-A and conventional LA in the following databases: MEDLINE, EMBASE, and the Cochrane library. Studies were reviewed independently and rated by Newcastle-Ottawa Quality Assessment Scale. The operative time, estimated blood loss in operation, the time to resume oral intake after surgery, postoperative hospital stay, and the visual analog pain scale (VAPS) score were included for analysis to compare the efficacy, while the complications together with the analgesia use were included for analysis to compare the safety. RESULTS: Nine studies with 171 LESS-A cases and 272 conventional LA cases were identified. Although operative time was longer in LESS-A (mean difference [MD] 15.46, 95% confidence interval [CI] 11.18 to 19.74), estimated blood loss (MD 4.72, 95% CI 12.08 to 21.52) and the time to resume oral intake (MD -0.04, 95% CI -0.19 to 0.11) were similar; LESS-A presented a shorter postoperative stay in hospital (MD -0.60, 95% CI -0.86 to -0.35) and lower VAPS score (MD -1.21, 95% CI -1.44 to -0.97). Besides, the risk of minor postoperative complications (risk ratio [RR] 1.74, 95% CI 0.78 to 3.87) was similar. The postoperative analgesia demand in total (RR 0.65, 95% CI 0.52 to 0.81) together with the analgesia usage lasting more than 24 hours after surgery (RR 0.35, 95% CI 0.21 to 0.58) were associated with lower risk in LESS-A, however. CONCLUSIONS: Based on current evidence, the operative time seems to be longer in LESS-A; however, operative blood loss and complications are similar. In addition, LESS-A presents a shorter hospital stay after surgery and more acceptable perception of pain than conventional LA.

The 1973 WHO Classification is more suitable than the 2004 WHO Classification for predicting prognosis in non-muscle-invasive bladder cancer.

BACKGROUND: Predicting the recurrence and progression of Non-muscle-invasive bladder cancer(NMIBC) is critical for urologist. Histological grade provides significant prognostic information, especially for prediction of progression. Currently, the 1973 and the 2004 WHO classification co-exist. Which system is better for predicting rumor recurrence and progression still a matter for debate. METHODOLOGY/PRINCIPAL FINDINGS: 348 patients diagnosed with Non-muscle invasive bladder cancer were enrolled in our retrospective study. Paraffin sections were assessed by an experienced urological pathologist according to both the 1973 and 2004 WHO classifications. Tumor recurrence and progression was followed-up in all patients. During follow-up, corresponding 5-year recurrence-free survival rates of G1, G2 and G3 were 82.1%, 55.9%, 32.1% and the 5-year progression-free survival rates were 95.9%, 84.4% and 43.3%, respectively. The 5-year recurrence-free survival rates of papillary urothelial neoplasm of low malignant potential (PUNLMP), low-grade papillary urothelial carcinoma(LGPUC) and high-grade papillary urothelial carcinoma (HGPUC) were 69.8%, 67.1% and 42.0% respectively and the 5-year progression-free survival rates were 100%, 90.9% and 54.8% respectively. In multivariate analysis, the 1973 WHO classification significantly associated with both tumor recurrence and progression(p=0.010 and p=0.022, respectively); the 2004 WHO classification correlated with tumor progression(p=0.019), while was not proved to be a variable that can predict the risk of recurrence(p=0.547). Kaplan-Meier plots showed that both the 1973 WHO and the 2004 WHO classifications were significantly associated with progression-free survival (p<0.0001, log-rank test). For prediction of recurrence, significant differences were observed between the tumor grades classified using the 1973 WHO grading system (p<0.0001, log-rank test), while a significant overlap was observed between PUNLMP and LG plots using the 2004 WHO grading system(p=0.616, log-rank test). CONCLUSION/SIGNIFICANCE: Both the 1973 WHO and the 2004 WHO Classifications are effective in predicting tumor progression in Non-muscle invasive bladder cancer, while the 1973 WHO Classification is more suitable for predicting tumor recurrence.

Periostin: a promising target of therapeutical intervention for prostate cancer.

BACKGROUND: In our recent study, Periostin was up-regulated in prostate cancer(PCa) compared with benign prostate hyperplasia (BPH) by proteomics analysis of prostate biopsies. We investigated the effect of sliencing Periostin by RNA interference (RNAi) on the proliferation and migration of PCa LNCap cell line. METHODS: All the prostate biopsies from PCa, BPH and BPH with local prostatic intraepithelial neoplasm(PIN) were analyzed by iTRAQ(Isobaric tags for relative and absolute quantification) technology. Western blotting and immunohistochemical staining were used to verify Periostin expression in the tissues of PCa. Periostin expression in different PCa cell lines was determined by immunofluorescence staining, western blotting and reverse transcription PCR(RT-PCR). The LNCap cells with Periostin expression were used for transfecting shRNA-Periostin lentiviral particles. The efficancy of transfecting shRNA lentiviral particles was evaluated by immunofluorescence, western blotting and Real-time PCR. The effect of silencing Periostin expression by RNAi on proliferation of LNCap cells was determined by MTT assay and tumor xenografts. The tissue slices from theses xenografts were analyzed by hematoxylin and eosin(HE) staining. The expression of Periostin in the xenografts was deteminned by Immunohistochemical staining and western blotting. The migration of LNCap cells after silencing Periostin gene expression were analyzed in vitro. RESULTS: Periostin as the protein of interest was shown 9.12 fold up-regulation in PCa compared with BPH. The overexpression of Periostin in the stroma of PCa was confirmed by western blotting and immunohistochemical staining. Periostin was only expressed in PCa LNCap cell line. Our results indicated that the transfection ratio was more than 90%. As was expected, both the protein level and mRNA level of Periostin in the stably expressing shRNA-Periostin LNCap cells were significantly reduced. The stably expressing shRNA-Periostin LNCap cells growed slowly in vitro and in vivo. The tissues of xenografts as PCa were verificated by HE staining. Additionally, the weak positive Periostin expressed tumor cells could be seen in the tissues of 6 xenografts from the group of down-regulated Periostin LNCap cells which had a significant decrease of the amount of Periostin compared to the other two group. Furthermore, our results demonstrated that sliencing Periostin could inhibit migration of LNCap cells in vitro. CONCLUSIONS: Our data indicates that Periostin as an up-regulated protein in PCa may be a promising target of therapeutical intervention for PCa in future.