ZNF492 and GPR149 methylation patterns as prognostic markers for clear cell renal cell carcinoma: Array­based DNA methylation profiling.

The present study aimed to identify novel methylation markers of clear cell renal cell carcinoma (ccRCC) using microarray methylation analysis and evaluate their prognostic relevance in patient samples. To identify cancer­specific methylated biomarkers, microarray profiling of ccRCC samples from our institute (n=12) and The Cancer Genome Atlas (TCGA) database (n=160) were utilized, and the prognostic relevance of candidate genes were investigated in another TCGA dataset (n=153). For validation, pyrosequencing analyses with ccRCC samples from our institute (n=164) and another (n=117) were performed and the potential clinical application of selected biomarkers was examined. We identified 22 CpG island loci that were commonly hypermethylated in ccRCC. Kaplan­Meier analysis of TCGA data indicated that only 4/22 loci were significantly associated with disease progression. In the internal validation set, Kaplan­Meier analysis revealed that hypermethylation of two loci, zinc finger protein 492 (ZNF492) and G protein­coupled receptor 149 (GPR149), was significantly associated with shorter time­to­progression. Multivariate Cox regression models revealed that hypermethylation of ZNF492 [hazard ratio (HR), 5.44; P=0.001] and GPR149 (HR, 7.07; P<0.001) may be independent predictors of tumor progression. Similarly, the methylation status of these two genes was significantly associated with poor outcomes in the independent external validation cohort. Collectively, the present study proposed that the novel methylation markers ZNF492 and GPR149 could be independent prognostic indicators in patients with ccRCC.

Surgical margin does not influence recurrence rate in pT1 clear cell renal cell carcinoma after partial nephrectomy: A multicenter study.

PURPOSE: To assess the risk factors of positive surgical margins (PSM) and the influence of margin status on recurrence in pT1 clear cell renal cell carcinoma (RCC) following partial nephrectomy (PN). MATERIALS AND METHODS: Patients (1,831) with pathologically confirmed stage T1 clear cell RCC were retrospectively analyzed following PN at eight institutions in Korea between 1999 and 2011. Demographics, operative data, pathological margin status, and site of recurrence were analyzed. RESULTS: Resection margins were positive in 31 patients (1.7% of the cohort) on final pathology. None of the clinicopathological parameters were significantly related to the marginal status (all P > 0.05). During a median follow-up of 32.5 months, local recurrences were observed in 0.4% of negative surgical margins. There was no local recurrence in any of the cases with PSM. Distant recurrences developed in 1.7% of negative surgical margins and 3.2% of PSM. There were no significant differences in recurrence-free survival by margin status (P = 0.566). CONCLUSIONS: Our multi-institutional data suggest that marginal status does not influence tumor recurrence risk in pT1 clear cell RCC after PN. Careful surveillance seems to be a sufficient strategy in this clinical scenario. J. Surg. Oncol. 2016;114:70-74. © 2016 Wiley Periodicals, Inc.

The c-MET Network as Novel Prognostic Marker for Predicting Bladder Cancer Patients with an Increased Risk of Developing Aggressive Disease.

Previous studies have shown that c-MET is overexpressed in cases of aggressive bladder cancer (BCa). Identification of crosstalk between c-MET and other RTKs such as AXL and PDGFR suggest that c-MET network genes (c-MET-AXL-PDGFR) may be clinically relevant to BCa. Here, we examine whether expression of c-MET network genes can be used to identify BCa patients at increased risk of developing aggressive disease. In vitro analysis, c-MET knockdown suppressed cell proliferation, invasion, and migration, and increased sensitivity to cisplatin-induced apoptosis. In addition, c-MET network gene (c-MET, AXL, and PDGFR) expression allowed discrimination of BCa tissues from normal control tissues and appeared to predict poor disease progression in non-muscle invasive BCa patients and poor overall survival in muscle invasive BCa patients. These results suggest that c-MET network gene expression is a novel prognostic marker for predicting which BCa patients have an increased risk of developing aggressive disease. These genes might be a useful marker for co-targeting therapy, and are expected to play an important role in improving both response to treatment and survival of BCa patients.

Clinical Implications and Prognostic Values of Prostate Cancer Susceptibility Candidate Methylation in Primary Nonmuscle Invasive Bladder Cancer.

DNA methylation is the most common and well-characterized epigenetic change in human cancer. Recently, an association between prostate cancer susceptibility candidate (PRAC) methylation and genitourinary cancer was proposed. The aim of the present study was to evaluate the association between PRAC methylation status and clinicopathological parameters and prognosis in long-term follow-up primary nonmuscle invasive bladder cancer (NMIBC). The clinical relevance of PRAC methylation was determined in 136 human bladder specimens (eight normal controls [NCs] and 128 primary NMIBCs) using quantitative pyrosequencing analysis. PRAC methylation was significantly higher in NMIBC patients than in NCs and was significantly associated with higher grade and more advanced stage of cancer. Kaplan-Meier estimates revealed significant difference in tumor recurrence and progression according to PRAC methylation status (both p < 0.05). Multivariate Cox regression analysis revealed that the PRAC methylation status was a strong predictor of recurrence (hazard ratio [HR], 2.652; p = 0.012) and progression (HR, 9.531; p = 0.035) of NMIBC. Enhanced methylation status of PRAC was positively associated with a high rate of recurrence and progression in NMIBC patients, suggesting that PRAC methylation may be a promising prognostic marker of NMIBC.

Impact of transobturator tape treatment on overactive bladder symptoms, particularly nocturia, in patients with mixed urinary incontinence.

PURPOSE: We assessed the impact of transobturator tape (TOT) treatment on overactive bladder (OAB) symptoms with a particular focus on nocturia in patients with mixed urinary incontinence (MUI). MATERIALS AND METHODS: In this retrospective cohort study, the medical records of 237 women who underwent TOT surgery for the treatment of MUI were reviewed. Of these, 86 patients (36.4%) had preoperative nocturia. Patients with neurological diseases or sleep disorders that could affect the voiding pattern were excluded. Patients who were being treated with anticholinergics and antidiuretic hormones were also excluded, which left 70 subjects eligible for analysis. Pre- and postoperative evaluations consisted of a physical examination, 3-day frequency-volume chart, and health-related quality of life questionnaires (King's health questionnaire, overactive bladder symptom score, and OAB-questionnaire). RESULTS: TOT treatment resulted in an overall significant improvement in OAB symptoms including nocturia. Frequency-volume charts revealed that TOT treatment significantly decreased the actual number of nightly voids (ANV) and the nocturnal bladder capacity index (NBCi) in the entire cohort. However, in a subgroup of women with nocturnal polyuria, there was no significant change in ANV or NBCi after the sling operation. Correlation analysis of the whole cohort revealed that the postoperative changes in NBCi correlated positively with postoperative changes in ANV. The nocturia-persisting group was more likely to have nocturnal polyuria and lower preoperative functional bladder capacity compared with the nocturia-improved group (p=0.024 and p=0.023, respectively). CONCLUSIONS: Our results demonstrated that the TOT procedure resulted in an overall significant improvement in OAB symptoms including OAB-related nocturia in patients who presented with MUI.

Hypertriglyceridemia and low high-density lipoprotein cholesterolemia are associated with increased hazard for urolithiasis.

PURPOSE: To assess the association between dyslipidemia and urolithiasis, a propensity score-matching study was performed. PATIENTS AND METHODS: Fasting blood samples were taken, and serum lipid profiles were measured in 655 stone formers (SF) and 1965 propensity score-matched controls between 2005 and 2011. The controls, from a health-screening program, did not have a history of dyslipidemia or statin use and have any evidence of stone disease, as determined by abdominal radiography, ultrasonography examination. Propensity score-matching with respect to age, sex, and body mass index was used to minimize selection bias, and the logistic regression analysis was adjusted for other components of metabolic syndrome. RESULTS: Compared with controls, the SF group had significantly higher mean triglyceride and lower total cholesterol, low-density lipoprotein (LDL) cholesterol, and high-density lipoprotein (HDL) cholesterol levels (each P<0.001). The SF group was also more likely to have hypertriglyceridemia and low HDL-cholesterolemia, and less likely to have hypercholesterolemia and high LDL cholesterolemia compared with controls (each P<0.05). When adjusted for other components of metabolic syndrome including obesity, presence of diabetes mellitus or hypertension, the odds ratio (OR) for urinary stones appeared with hypercholesterolemia (OR=0.747, P=0.003), hypertriglyceridemia (OR=1.901, P<0.001), low HDL cholesterolemia (OR=1.886, P<0.001) and high LDL cholesterolemia (OR=0.610, P<0.001). CONCLUSIONS: Our study implies that dyslipidemia may play a crucial part in urinary stone risk.

Natural history of asymptomatic renal stones and prediction of stone related events.

PURPOSE: The appropriate management for asymptomatic renal stones remains unclear. We assessed the natural history and progression rate of such stones and identified clinical factors associated with an increased risk of stone related events. MATERIALS AND METHODS: We retrospectively reviewed the medical records of 201 male and 146 female patients with asymptomatic renal stones. It was recommended that patients be followed every 6 months. Mean followup was 31 months (range 6 to 180). Patients were divided into 2 groups by stone related events, including spontaneous stone passage, flank pain, stone growth or the need for intervention during followup. RESULTS: Spontaneous passage occurred in 101 patients (29.1%). Of the patients 186 (53.6%) and 161 (46.4%) did and did not have stone related events, respectively. Of the whole cohort 85 patients (24.5%) required intervention but only 4.6% needed surgery. At 19 months after diagnosis 50% of the patients had a symptom. Those with stone related events were more likely to be younger (mean ± SD age 46.6 ± 12.7 vs 49.3 ± 12.6 years) and male, and have a stone history (p = 0.047, 0.017 and 0.014, respectively). Male gender significantly decreased the probability of freedom from stone related events (log rank test p = 0.0135) and it was an independent predictor of stone related events (HR 1.521, p = 0.009). Younger patients, and those with smaller stones and no stone growth were more likely to experience spontaneous passage and less likely to undergo intervention (each p <0.05). CONCLUSIONS: Asymptomatic renal stones can be followed safely but long-term followup is necessary. Periodic followup and early intervention should be recommended in patients with risk factors.

Clinical evaluation of the depigmenting effect of Glechoma Hederacea extract by topical treatment for 8 weeks on UV-induced pigmentation in Asian skin.

UV is a major environmental factor inducing and worsening the symptoms of hyperpigmentation disorders such as freckles, melasma and solar lentigines. During UV-induced skin inflammatory reactions, pro-inflammatory mediators initiate the production of various paracrine melanogenic factors (α-MSH, SCF, ET-1, bFGF and NO) in keratinocytes. These paracrine factors activate melanin synthase in melanocytes through the paracrine network between melanocytes and keratinocytes. Glechoma hederacea (GH) is a herbal plant used in oriental medicine to treat inflammation. Its anti-inflammatory effects, through inhibition of NO synthesis (NOS) as well as the pro-inflammatory cytokine TNF-α, have been reported. However, there has not yet been any report of a depigmenting effect. In this study of placebo-controlled, 8 week topical treatment with a 1% GH extract lotion on UV-induced pigmented spots in female Asian subjects, significant effects of anti-inflammation and depigmenting were proven. The depigmenting effect of GH seems to be related to inhibiting the secretion of pro-inflammatory cytokines and melanogenic paracrine factors from keratinocytes, rather than to direct inhibition of melanogenic activities in melanocytes.

A global gene expression analysis of the peripheral blood mononuclear cells reveals the gene expression signature in psoriasis.

BACKGROUND: Psoriasis is a chronic inflammatory skin disease that affects approximately 1~3% of the general population. OBJECTIVE: We performed cDNA microarray analysis with using the dendrimer labelling method to investigate the gene expression profile in the peripheral blood mononuclear cells (PBMCs) of psoriatic patients. METHODS: The peripheral blood mononuclear cells of 5 patients with psoriasis and 8 control subjects were used in the gene expression analyses of psoriasis. RESULTS: We identified 212 differentially expressed genes that showed at least a two-fold induction and/or reduction in psoriatic patients. Among those, 63 genes, including CD44, CD56 and IL7R, were induced, while 139 genes, including the sphingosine kinase 1 and p16-INK genes, were reduced in the psoriatic patients. CONCLUSION: We can speculate that these genes may have a role for the pathogenesis of psoriasis via their affecting different cellular functions. Our results suggest a possible mechanism by which activated immune cells migrate from the blood to the skin in psoriatic patients, and we provide novel putative targets for developing drugs to treat psoriasis.

Expression of neutrophil gelatinase-associated lipocalin in calcium-induced keratinocyte differentiation.

In a previous search for the differentially expressed genes in keratinocyte differentiation, we identified neutrophil gelatinase-associated lipocalin (NGAL) as a calcium-induced gene. In this study, we further verified the expression of NGAL in cultured keratinocytes as well as in several skin diseases. Reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and ELISA clearly showed that NGAL expression was markedly increased in calcium-induced keratinocyte differentiation in vitro. However, in our previous report, NGAL expression was not detected in normal skin tissue except for hair follicle by in situ hybridization and immunohistochemistry, indicating the difference of cell status between in vitro and in vitro conditions. Interestingly, NGAL expression was highly increased in psoriasis-like inflammatory disorders (lichen planus and pityriasis rubura pilaris) and skin cancers (keratoacanthoma and squamous cell carcinoma), implying that NGAL may be related with the epidermal hyperplasia. Collectively, these results reveal the potential importance of NGAL in the maintenance of skin homeostasis.

IFN-gamma regulates the expression of B7-H1 in dermal fibroblast cells.

BACKGROUND: Programmed cell death ligand 1 (B7-H1) was recently cloned in antigen presenting cells (APCs) and represents a third member of the B7 family. Thus, B7-H1 may be a novel target for clinical intervention in human inflammatory disease. OBJECTIVE: The aim of this study is to investigate the signal transduction mechanism and transcriptional regulation of B7-H1 expression in human dermal fibroblasts. METHODS: We performed reverse transcription PCR (RT-PCR) for the detection of mRNA expression, luciferase reporter assays with B7-H1 promoter constructs, and Western blot analysis. RESULTS: From RT-PCR analysis, IFN-gamma can induce the expression of B7-H1 mRNA in dermal fibroblast. This expression is similar to the results of luciferase reporter assay with B7-H1 promoter. Western blot analysis and EMSA revealed that NF-kappaB transcription factors mediate the induction of B7-H1 expression via the transient phosphorylation of ERK1/2 and PI3K when cells are stimulated by IFN-gamma. Also, Specific destruction of the NF-kappaB binding site abolished the induction of the promoter activity by IFN-gamma. CONCLUSION: Our data not only provides the first evidence to demonstrate that dermal fibroblast express the B7-H1 mRNA in the process of skin inflammation, but also suggests the involvement of NF-kappaB and MAPK and PI3K, that may play some important roles in inflammation process in human skin diseases.

Suppression of the onset and progression of collagen-induced arthritis by chebulagic acid screened from a natural product library.

OBJECTIVE: Chebulagic acid (CHE) from the immature seeds of Terminalia chebula was identified from a natural product library as a potent suppressor of T cell activity. This study examined the effectiveness of CHE against the onset and progression of collagen-induced arthritis (CIA) in mice. METHODS: Arthritis was induced in DBA/1J mice by subcutaneous immunization with bovine type II collagen on days 0 and 21. CHE was administered intraperitoneally for 3 weeks, either as prophylaxis (10 or 20 mg/kg) before disease onset or as therapy (20 mg/kg) after disease onset. Clinical scores, serum antibody levels, and cytokines were measured, and flow cytometric analysis and real-time reverse transcription-polymerase chain reaction were performed to evaluate the knee joints of mice with CIA. RESULTS: In both the prophylactic and therapeutic CHE dosing models, all clinical scores, serum levels of total and anticollagen IgG, and levels of interleukin-10 (IL-10) and IL-6 were reduced, while serum levels of transforming growth factor beta (TGFbeta) were markedly elevated. The number of granulocytes was reduced, but the proportion of CD4+,CD25+ T cells was greater in the knee joints of CHE-treated CIA mice. Expression of Foxp3 and TGFbeta messenger RNA was also augmented significantly in the knee joints of CHE-treated CIA mice in the therapeutic dosing model. CONCLUSION: CHE significantly suppressed the onset and progression of CIA in mice. Immune suppression via the induction of TGFbeta and CD4+,CD25+ T cells may represent a new strategy in the development of therapies for managing rheumatoid arthritis and other inflammatory diseases.