Takuya Yamanobe

Deterioration of ischemia/reperfusion-induced acute renal failure in SOD1-deficient mice

Reactive oxygen species (ROS) are likely candidates for involvement in ischemia/reperfusion-induced acute renal failure (ARF). In this study, the issue of whether superoxide dismutase (SOD1)-deficiency exacerbates the ischemia/reperfusion-induced ARF was examined. At two weeks after a right nephrectomy of mice, the left renal vessels were clipped to induce renal ischemia and were then released after 45 min. The severe renal damage observed at one day was partially recovered at seven days after the induction of ischemia. SOD1− / − mice suffer from severe ARF compared with SOD1+/ − and SOD1+/+ mice. The damage was more evident in aged animals (24–28 week old) than younger ones (10–12 week old). The expression of major antioxidative and redox enzymes, except for CuZnSOD, were substantially unchanged. Thus, the increased ARF in SOD1− / − mice appears to be mainly attributable to a deficiency in CuZnSOD. These data support the view that ROS are exacerbating factors in ischemia/reperfusion-induced ARF.

Aggravation of ischemia-reperfusion-triggered acute renal failure in xCT-deficient mice.

This study examined the question of whether deficiency of xCT, a cystine-transporter gene, exacerbates ischemia-reperfusion-induced acute renal failure (ARF). Two weeks after the right nephrectomy of male mice at 16-18weeks of age, the left renal vessels were clamped for 45min to induce renal ischemia. After (24h) induction of ischemia, xCT(-/-) mice had elevated concentrations of blood urea nitrogen and creatinine indicative of ARF, while in xCT(+/-) and xCT(+/+) mice, these parameters did not differ from the sham-operated mice. Immunohistochemical analyses of kidneys using antibodies against the oxidative stress markers revealed stronger staining in xCT(-/-) mice compared with xCT(+/+) mice. Induction of xCT mRNA in the kidneys of xCT(+/+) mice was demonstrated using reverse transcriptase (RT)-PCR analysis and was further confirmed using quantitative RT-PCR. These data provide the first in vivo evidence that xCT is induced by oxidative stress and helps prevent ischemia-reperfusion injury to kidneys.

The Outcome of Prostate Cancer Screening in a Normal Japanese Population with PSA of 2–4 ng/ml and the Free/Total PSA Under 12%

OBJECTIVE No previous study has reported the numbers of prostate cancer (PC) patients existing among a normal Japanese population with prostate-specific antigen (PSA) < 4 ng/ml. The aim of this study was to elucidate the performance of %free PSA as a screening tool for a normal Japanese population with PSA of 2-4 ng/ml and to examine the characteristics of cancer detected using this criterion. METHODS We conducted a prospective, multi-center study to evaluate the performance of %free PSA among a normal Japanese population. We decided on a %free PSA cutoff value of 12% according to the preliminary results. A total of 5548 consecutive screening volunteers aged 50-79 years were enrolled in the project. Men with total PSA > 4 ng/ml, or men with total PSA of 2-4 ng/ml and %free PSA of < or =12% were indicated to undergo 12 core biopsies. RESULTS There were 826 (14.9%) men with PSA of 2-4 ng/ml. Among them, those with %free PSA of < or =12% numbered 100 (12.1%). Forty-nine out of 100 men (49%) received biopsy, and 16 PC patients were detected. Among 10 patients undergoing radical prostatectomy, seven were associated with extra-prostatic extension (pT3) or high-grade cancer (Gleason score > or = 8). CONCLUSIONS We confirmed the ability of %free PSA and demonstrated that there are considerable numbers of PC patients among the normal Japanese population with PSA of 2-4 ng/ml. We ascertained that cancers detected in this study had a variety of tumor characteristics, including those of an aggressive nature.

MP88-18 ERK5 IS A PROMISING THERAPEUTIC TARGET FOR CLEAR CELL RENAL CELL CARCINOMA

RASAL2 methylation or c-FOS mRNA or VEGFA mRNA in RCC tissues. Overexpression of RASAL2 in 786-O cells could inhibit the recruitment and tube formation of HUVECs, while RASAL2 knockdown (KD) in ACHN cells enhanced the recruitment and tube formation of HUVECs in vitro. Also, overexpression of RASAL2 could inhibit tumorigenecity of xenografts. Mechanistically, RASAL2 KD could enhance the phosphorylation of GSK3 and upregulate the expression of c-FOS and VEGFA. Furthermore, RASAL2 was inversely correlated with VEGFA and CD31 in tissues from human RCC specimens and xenografts. CONCLUSIONS: RASAL2 was downregulated in RCC tissues, which could lead to tumor angiogenesis via p-GSK3/c-FOS/VEGFA signaling pathway. Therefore, RASAL2 could be a potential target to prevent patients with RCC from resistance to anti-vascular therapy.

Successful Treatment with Paclitaxel, Carboplatin, and Gemcitabine as Second-line Chemotherapy for Recurrent Urothelial Carcinoma of the Bladder with Glandular Differentiation After Radical Cystectomy: A Case Report

Urothelial carcinoma of the bladder (UCB) with glandular differentiation is a histological variant (HV) that is more likely to have positive extravesical tumors or nodes than those in pure UCB. Cisplatin-based neoadjuvant chemotherapy (NAC) prior to radical cystectomy (RC) is more effective for pure UCB; however, few reports are available on second-line chemotherapy for recurrence of UCB with HV. Here we report a 65-year-old Japanese male diagnosed with local recurrence UCB with HV after NAC + RC who safely achieved complete response with paclitaxel, carboplatin, and gemcitabine combination chemotherapy.

SAFETY AND EFFICACY OF CONTINUOUS ADMINISTRATION OF ANTITHROMBOTIC DRUGS DURING TRANSURETHRAL RESECTION OF BLADDER TUMORS

(Objectives) We evaluated the safety and efficacy of continuous administration of antithrombotic drugs during transurethral resection of bladder tumors (TURBT). (Methods) We performed a retrospective review of 351 TURBT procedures performed at our institute from April 2011 to October 2015. Among these, antithrombotic drugs were continued in 31 TURBT cases throughout their perioperative period (continuation group), antithrombotic drugs were discontinued in 26 TURBT cases (discontinuation group), and bridging anticoagulation therapy with heparin after interruption of antithrombotic drugs was performed in 4 TURBT cases (heparin bridging group). The safety and efficacy of continuous administration of antithrombotic drugs during TURBT was evaluated by comparing the rate of perioperative complications, median operative time, duration of hematuria, urethral catheter placement, duration of stay after surgery, and by comparing the duration of stay among the three groups. (Results) The median operative time was 40.0 min in the continuation group, 39.0 min in the discontinuation group, and 31.0 min in the heparin bridging group with no significant differences. There were no significant differences in the median duration of hematuria (1.00 days vs. 1.00 days vs. 1.00 days), urethral catheter placement (3.00 days vs. 2.50 days vs. 2.00 days), or stay after TURBT (4.00 days vs. 3.50 days vs. 3.00 days) among the continuation, discontinuation, and heparin bridging groups. The median duration of stay was 6.00 days in the continuation group, 7.00 days in the discontinuation group, and 16.0 days in the heparin bridging group with significant differences between the continuation group vs. the heparin bridging group and the discontinuation group vs. the heparin bridging group. The rate of complications was 6 (19.4%) in the continuation group and 3 (11.5%) in the discontinuation group with no significant differences. However, a serious complication, cerebral infarction leading to hemiplegia, occurred in the discontinuation group. (Conclusion) Continuous administration of antithrombotic drugs during TURBT is considered to be safe and useful because it may prevent serious thromboembolism without adversely affecting the perioperative course.

SMAC-N7 PEPTIDE WAS MORE POTENT THAN XIAP GENE TARGETING IN SENSITIZING RENAL CANCER CELLS TO APOPTOTIC STIMULI

253 SMAC-N7 PEPTIDE WAS MORE POTENT THAN XIAP GENE TARGETING IN SENSITIZING RENAL CANCER CELLS TO APOPTOTIC STIMULI Tomoyuki Kato*, Sei Naito, Hisashi Kawazoe, Tomohiro Shibasaki, Yuko Nakano, Takuya Yamanobe, Akira Kajinuma, Akinori Muto, Bilim Vladimir, Akira Nagaoka, Yoshihiko Tomita. Yamagata, Japan. INTRODUCTION AND OBJECTIVE: Renal cell carcinoma (RCC) is known to be resistant to chemoand radiotherapy due to a high apoptotic threshold. Inducing apoptosis in RCC cells leads to tumor regression and a molecule responsible for avoiding apoptosis can be a biomarker of molecular target therapy. In this study we tried to manipulate function of the strong anti-apoptotic molecule, XIAP, using RNAi and a peptide mimicking its inhibitory molecule, the second mitochondriaderived activator of caspase (Smac), releasing from mitochondria when a cell is exposed to apoptotic stimuli. METHODS: The surgical specimens from 34 patients who underwent surgery for RCC from January to December 2005 at the Yamagata University Hospital were subjected to immunohistochemical staining with anti-Smac and XIAP antibodies. Staining score was calculcuated by staining intensity multiplied by percent of positive cells, range 0-300. Caki1 RCC cell line with high levels of XIAP was transfected with XIAP targeting shRNA vector and stable clones were generated. The cell permeable peptide Smac-N7 was applied to in vitro treatment of the Caki1. Detection of apoptosis was performed by Hoechst 33342 and Giemsa staining and cell viability was checked by MTS assay. RESULTS: Smac was found in all tumors with staining scores varying from 200 to 300 and staining intensity was weaker than in normal kidneys. XIAP expression levels increased from pT1 (mean score 164.4±108.2) to pT2(195.0±83.3) and pT3(266.7±57.7), as well as from grade 1 (75.0± 98.7) to grade 2 (201.2 ± 93.6) (p<0.05). XIAP was knocked down by RNAi in clone #14 by 81.6% and in clone #19 by 85.3%. Compared to the parental and mock-transfected cells, neither clone was more sensitive to conventional chemotherapeutic agents, but both clones were more susceptible to Fas-stimulation (p<0.0001) and to pharmacological Bcl-2 inhibition (p<0.0001), as well as to a combination of the two (p<0.0001). Exposure of Caki1 cells to Smac-N7 peptide