Minekatsu Taga

Expression of MicroRNAs associated with Gleason grading system in prostate cancer: miR-182-5p is a useful marker for high grade prostate cancer

Expression profiles of some microRNAs (miRNAs) were associated with clinicopathological findings in human prostate cancer (PC), but the relative expression of miRNAs among Gleason patterns (GPs) remains unclear. In this study, we investigated the expression of several known microRNAs in each GP of PC.

A potential role for 6-sulfo sialyl Lewis X in metastasis of bladder urothelial carcinoma

OBJECTIVES It is widely accepted that sialyl Lewis X (sLeX) and sialyl Lewis A (sLeA, also known as CA 19-9) glycans expressed on cancer cells function in E-selectin-mediated metastasis. Recently, it was reported that 6-sulfo sLeX glycans detected by the MECA-79 monoclonal antibody are expressed in roughly a quarter of gastric adenocarcinoma cases, and that these cases show a poorer prognosis than MECA-79-negative cases do. The present study was undertaken to assess expression of 6-sulfo sLeX glycans in bladder urothelial carcinoma and evaluate potential clinical implications. MATERIALS AND METHODS We analyzed 78 specimens representing bladder urothelial carcinoma, as well as 4 bladder urothelial carcinoma cell lines, by immunostaining with a battery of anticarbohydrate antibodies. We also undertook an E-selectin·IgM chimera binding assay to assess E-selectin binding to 6-sulfo sLeX expressed on bladder urothelial carcinoma cells and performed reverse transcription polymerase chain reaction and complementary DNA transfection to determine which N-acetylglucosamine-6-O-sulfotransferases function in 6-sulfo sLeX biosynthesis in those cells. Finally, we performed double-immunofluorescence staining for MECA-79 and either CD3 or CD8 to evaluate potential association between high endothelial venule (HEV)-like vessels and tumor-infiltrating T lymphocytes. RESULTS 6-Sulfo sLeX glycans were expressed in ~20% of bladder urothelial carcinoma cases, particularly in plasmacytoid and micropapillary variants. Positive cells were also bound by E-selectin·IgM chimeras in a calcium-dependent manner. Transcripts encoding N-acetylglucosamine-6-O-sulfotransferase-2 were detected preferentially in HT-1197 bladder urothelial carcinoma cells expressing 6-sulfo sLeX, and transfection of the enzyme complementary DNA into HT-1376 cells, which do not express 6-sulfo sLeX glycans, resulted in cell surface expression of 6-sulfo sLeX. Furthermore, 6-sulfo sLeX glycans were expressed in HEV-like vessels induced in and around lymphocyte aggregates formed near carcinoma cell nests. These HEV-like vessel-associated tumor-infiltrating lymphocytes were composed primarily of CD3(+) T cells, with a fraction of CD8(+) cytotoxic T cells. CONCLUSIONS Our findings indicate that 6-sulfo sLeX glycans likely play 2 roles in bladder urothelial carcinoma progression: one in lymphocyte recruitment to enhance antitumor immune responses, and the other in E-selectin-mediated tumor cell adhesion to vascular endothelial cells, which is potentially associated with metastasis.

IPSS is lower in hypertensive patients treated with angiotensin-II receptor blocker: Posthoc analyses of a lower urinary tract symptoms population†‡

The existence of Angiotensin‐II (Ang‐II) receptors in the bladder wall and the pronounced contractile effect of Ang‐II on the human detrusor muscle have been well established. Studies have presented the role of Ang‐II as a mediator in smooth muscle growth and collagen production in the bladder with outlet obstruction. We investigated the associations between male lower urinary tract symptoms (LUTS) and hypertension (HT), and examined whether the medications used for HT treatment, particularly Ang‐II receptor blockers (ARBs) influence LUTS.

Castration increases PGE 2 release from the bladder epithelium in male rats

Aims: Androgen deprivation therapy has been widely used for the treatment of prostate cancer. While sexual side effects including decreased sexual desire and function are well studied, there are only limited reports about its influences on lower urinary tract symptoms. The aim of this study is to clarify the influences of castration in male rats. Methods: Ten‐week‐old male rats were divided into treatment group (bilateral orchiectomy) and control group (sham surgery). Two‐months after the surgery, adenosine triphosphate (ATP), prostaglandin E2 (PGE2), and nerve growth factor (NGF) released from stretched bladder epithelium were measured by luciferin‐luciferase assay or ELISA. The mRNA levels of bladder cyclooxygenase‐1 (COX‐1) and cyclooxygenase‐2 (COX‐2) were determined by real‐time PCR. The protein level of bladder COX‐2 was analyzed by western blot analysis. Bio‐Plex Pro cytokine assay was performed to quantify the level of proinflammatory cytokine interleukin (IL)‐1&bgr; in the bladder. Results: The PGE2 release from stretched bladder epithelium was significantly increased after castration, which increased more than 50% compared with control. On the other hand, those of ATP and NGF were not different from those of the controls. Testosterone replacement restored the PGE2 increase. Castration significantly increased bladder IL‐1&bgr; protein level and COX‐2 at both mRNA and protein levels, whereas caused no marked changes in the COX‐1 mRNA level. Conclusions: These findings suggest that castration induces inflammation in the rat bladder, which causes elevated PGE2 release from bladder epithelium and may finally contribute to the disruption of bladder storage function.

Prostatic stromal inflammation is associated with bladder outlet obstruction in patients with benign prostatic hyperplasia

Benign prostatic hyperplasia (BPH) is a common urologic disease in older men. Prostatic inflammation research has focused on the magnitude of inflammation; its location has received little attention. We investigated whether the anatomic location of prostatic inflammation is related to the severity of lower urinary tract symptoms (LUTS), measured subjectively and objectively.

Clinicopathological implications to micropapillary bladder urothelial carcinoma of the presence of sialyl Lewis X-decorated mucin 1 in stroma-facing membranes

OBJECTIVES Bladder urothelial carcinoma (UC) comprises more than 90% of all bladder cancers. Among several UC variants, micropapillary UC (MPUC) is a rare one with high potential for lymphovascular invasion and subsequent lymph node metastasis. Histologically, MPUC is characterized by the presence of small papillary carcinoma cell clusters surrounded by lacunar spaces. Immunohistochemically, the outer circumference of these clusters, that is, the stroma-facing membrane of carcinoma cells, is reportedly almost invariably positive for mucin 1 (MUC1) protein and to a lesser extent for sialyl Lewis X (sLeX) carbohydrates; however, the clinicopathological implications of these expression patterns have not been fully investigated. MATERIALS AND METHODS We performed immunohistochemical analysis of MPUC (n = 11) and conventional UC (n = 57) for MUC1 and sLeX to determine whether these factors immunolocalized. Dual immunofluorescence staining was also carried out to assess MUC1 and sLeX colocalization. We also performed Western blot analysis of Chinese hamster ovary cells misexpressing both recombinant epitope-tagged MUC1 and glycosyltransferases enabling sLeX biosynthesis. RESULTS MPUC samples preferentially exhibited both MUC1 protein and sLeX carbohydrate expression on the stroma-facing membrane of carcinoma cells. Based on univariate analysis, MUC1 expression in that pattern was positively correlated with tumor extension, lymphovascular invasion, lymph node metastasis, disease stage, and relatively poor patient prognosis. A comparable sLeX expression pattern also correlated positively with tumor extension and nodal metastasis. Based on multivariate analysis, localization of MUC1 and sLeX on the stroma-facing side of the membrane was positively correlated with lymph node metastasis. CONCLUSIONS Overall, our immunofluorescence findings as well as immunoprecipitation analyses of Chinese hamster ovary cell transfectants strongly suggest that MUC1 is a potential scaffold protein for sLeX carbohydrates in MPUC. Both MUC1 and sLeX may cooperatively contribute to MPUC histogenesis and clinicopathological characteristics.

MP29-12 PRECLINICAL ASSESSMENT OF EARLY THERAPEUTIC EFFECT OF SUNITINIB ON RENAL CELL CARCINOMA USING 18F-FLUOROTHYMIDINE POSITRON EMISSION TOMOGRAPHY IN A XENOGRAFT MODEL

vaccines. Clinical and immunologic results vary. We have recently shown that genomic profiling of peripheral blood mononuclear cells (PBMC) may predict response. Transcription factors play a key role in the myelopoesis of monocytes and DC. We hypothesized that the profile of key transcription factors plays a key role in the efficacy of DC vaccines. METHODS: Monocytes from RCC patients and healthy donors were isolated via cold agglutination or ELUTRA. For some experiments, DC were matured with GM-CSF/ IL-4 and a variety of maturation protocols (TNF-alpha, cytokine cocktail, Flt-3).RNA was extracted using QIAGEN RNeasy Mini Kit. It’s concentration and quality was assessed using Nanodrop (Thermo Scientific) and Bioanalyzer (Agilent Technologies). Only high quality RNA (RIN > 7.2) was used for reverse transcription (Bio-Rad iScript cDNA synthesis kit) and real time qPCR (in triplicates using ABI SYBR-Select Master Mix with target specific primers and the ABI 7900HT Thermocycler). The expression of key transcription factors IRF-4, IRF-8, IL-10, IL-12 were analyzed using previously reported primer pairs. To determine relative concentrations, cycle threshold (Ct) values were compared to the reference gene GAP-DH. The statistical analysis was conducted in JMP 11 from SAS. RESULTS: Healthy donors demonstrated a significant variation of expression levels of all transcription factors examined. When DC were matured with a variety of maturation protocols, these resulted in significant differences in expression of all transcription factors. The greatest variations were observed in the expression levels of IRF-4, IL10 and IL-12. The greatest induction of IRF-4 and IL-12 and lowest induction of IL-10 were observed with the cytokine cocktail. In an IFNgamma release assay, these cells also demonstrated the greatest T cell stimulatory capacity. RCC patient monocytes showed showed significantly lower expression of IRF-4 and IRF-8 than healthy age-adjusted donor monocytes (p1⁄40.004 and 0.0214, respectively). Immunotherapy resulted in upregulation of IL-10 when compared to pretreatment mRNA levels (p1⁄40.007). CONCLUSIONS: In summary, this is the first study to assess the profile of key transcription factors as predictors for DC vaccine quality. We demonstrate significant inter-individual variability in monocytes prior to DC vaccine and throughout the generation of DC vaccines. These data may lead to improved cellular anti-cancer vaccines.

Surgical Treatment of Adrenal Gland Metastasis Originating from Small Cell Carcinoma of the Urinary Bladder

We report a rare case of a solitary adrenal metastasis from small cell carcinoma of the urinary bladder that was successfully treated with surgical resection. A 71-year-old man was suffering from bladder tamponade for hematuria. Computed tomography (CT) revealed a bladder tumor at the left wall. The patients underwent radical cystectomy. Histopathological results were obtained in small cell carcinoma of the bladder with muscle invasion. Thus, he received two courses of adjuvant etoposide and cisplatin chemotherapy, followed by the regimen for small cell lung cancer. Seven months after surgery, follow-up CT showed a gradually enlarged mass enhanced heterogeneously in the right adrenal gland. There was a solitary adrenal metastasis without any other metastasis; therefore, we performed right laparoscopic adrenalectomy. The patient has remained uneventful for four years after the adrenal gland surgery. For patients who have a solitary adrenal metastasis, adrenalectomy may provide a survival benefit.

Abstract 5033: Relationship between microRNA expression and Gleason grading system in prostate cancer

Background: Recent studies have shown that some microRNA (miRNA) expression profiles are associated with clinicopathological findings, including “Gleason score (GS)” in human prostate cancer (PC). However, the direct relationship between “Gleason pattern (GP)” and miRNA expression remains unclear. In this study, we investigated the miRNA expression profile in each GP (GP 3, GP 4, and GP 5). Methods: Formalin-fixed, paraffin-embedded (FFPE) tissue samples obtained from radical prostatectomy (patient set 1, n = 43, including [GP 3] n = 22, [GP 4] n = 35, and [GP 5] n = 12) and prostate needle biopsy (patient set 2, n = 9, [GP 4] n = 9) were used. Cancer tissues and adjacent normal counterparts were collected separately using laser-captured microdissection with subsequent isolation of total RNA. Real time reverse transcription polymerase chain reaction (RT-PCR) was performed to determine the relative expression of miRNAs, including miR-31, -34c, -96, -182, -183, -205, -221, and miR-222, which are currently reported to be involved in the progression of PC. Results: In the radical prostatectomy samples (patient set 1), relative expression (mean ± SE) of miR-31, miR-34c, and miR-205 in every GP was significantly decreased (miR-31: [GP 3] 0.028 ± 0.007, [GP 4] 0.047 ± 0.017, and [GP 5] 0.071 ± 0.022; miR-34c: 0.208 ± 0.066, 0.419 ± 0.219, and 0.210 ± 0.054; and miR-205: 0.021 ± 0.008, 0.015 ± 0.003, and 0.050 ± 0.023, all p-values Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5033. doi:1538-7445.AM2012-5033