e426 THE JOURNAL OF UROLOGY� Vol. 191, No. 4S, Supplement, Sunday, May 18, 2014

Bladder Cancer: Basic Research IV

Moderated Poster

Sunday, May 18, 2014 3:30 PM-5:30 PM

MP39-01CALCIUM MEDIATED AUTOPHAGY INDUCTION PRESERVECELL SURVIVAL DURING ENDOPLASMIC RETICULUM STRESSIN BLADDER CANCER

Rani Ojha, Shrawan Singh*, Arup Mandal, Shalmoli Bhattacharyya,Vivekanand Jha, Chandigarh, India

INTRODUCTION AND OBJECTIVES: Anticancer agents havebeen shown to induce endoplasmic reticulum (ER) stress and autophagyin various cancer cells. Autophagy acts as an adaptive mechanism forcancer cell survival. However, the mechanism and the effect of ER stressinduced autophagy in urothelial cancer cell survival is not well understood.

METHODS: Materials consisted of high and low grade urothelialcancer (UC) tissue and normal urothelium. ER stress was induced bytunicamycin and thapsigargin and measured by expression of various ERstress marker i.e. Grp-78, Ero-1La, PDI, IRE1a and PERK. ER stress in-duction was also validated by the measurement of Ca+2, IP3 and DAGassay. Autophagy was studied by lysotracker and LC3 immuno-staining.ExpressionofAMP-activatedproteinkinase (AMPK)andmammalian targetof rapamycin (mTOR) was also studied for determination of autophagy.ER stress induced autophagy on cell survival was studied in UC derivedprimary culture cells and T24 bladder cancer cell line, by the measurementof cell viability,mitochondrialmembrane potential, and caspases activation.

RESULTS: Treatment of tunicamycin and thapsigargin inducedthe expression of ER stress marker at six hours and increased up-to 12hours in UC cells as well as in normal urothelium (Fig.1A). There wasincreased Ca+2 flux, IP3 and DAG activity. Autophagosomes alsoincreased in number,more so in highgradeUC (Fig.1B). Theexpressionofphophorylated-AMPK increased and the expression of phosphorylated-mTOR decreased. Addition of autophagy inhibitor, wortmannin (Wm)along with tunicamycin or thapsigargin induced cell death in both gradeof UC (Fig.2A). Treatment of calcium chelater (EGTA) or pan caspaseinhibitor, z-VAD-fmkblocked the cell death induced byWm(Fig.1C). Therewas increased caspase-12, -9 and -3 expression (Fig.1D).

CONCLUSIONS: Ca+2 serves as a potent inducer of autophagyduring ER stress in UC as well as in normal urothelium. Ca+2 mediatedautophagy is regulated by IP3-AMPK-mTOR pathways and inhibition ofCa+2 mediated autophagy leads to caspase mediated intrinsic apoptoticcell death, suggesting that autophagy plays important roles in cell survival.Further elucidation of the molecular mechanisms linking ER stress toautophagy is needed for therapeutic intervention in urothelial carcinoma.

Source of Funding: Indian Council of Medical Research, New Delhi

MP39-02CORE MUSCLE SIZE PREDICTS 6-MONTH MORTALITYIN PATIENTS UNDERGOING RADICAL CYSTECTOMY

Hailey Allen, E Jason Abel, Daniel D. Shapiro, Fangfang Shi,Aaron Potretzke, David F. Jarrard, Timothy J. Ziemiewicz,Tracy M. Downs*, Madison, WI

INTRODUCTION AND OBJECTIVES: Loss of skeletal musclemass, termed sarcopenia, is a reproducible marker of overall frailty andis closely linked to surgical outcomes. Core muscle size seems torepresent a unique domain of preoperative surgical risk assessment toidentify patient’s who may not benefit from curative oncologic surgery.The objective of this study was to evaluate the ability of total psoasmuscle area (TPA) to predict mortality in patients undergoing radicalcystectomy (RC).

METHODS: Preoperative computed tomographic (CT) studieswere retrospectively analyzed in 128 of the 145 patients who underwentRC for urothelial carcinoma between January 2009 and May 2013.Cross-sectional area in mm2 was measured of the left and right psoasmuscles at the level of the L4 transverse processes. The left and rightpsoas muscle areas were summed to obtain a total psoas muscle area(TPA). Patients were divided into tertiles of TPA, with the high TPAgroup serving as the reference group for the analysis. Outcomesmeasured included 6-month mortality.

RESULTS: 98 patients (76.5%) were male and 30 patients(23.4%) were female. Median follow-up 25.4 months. Median age was68 years old (IQR 58-76.5), median BMI 28.1 (IQR 24.8-30.9), andmedian preoperative albumin was 3.7 g/dl (IQR 3.5-4.2). Median TPAwas 2321.5 mm2 (IQR 1908.6-2785.2) On univariate analysis overallsurvival was significantly worse for the low TPA group compared to thehigh TPA group - HR 2.85 (CI 1.4-5.7).

6-month mortality (p¼0.01) rates were significantly higher in the lowTPA group compared to the high TPA group.

CONCLUSIONS: Additional measures to improve preopera-tive risk assessment are necessary for patients undergoing radicalcystectomy. Core muscle size, an objective measure of frailty wascorrelated with early mortality following RC in this study. Multi-center studies in larger cohorts are necessary to valid thesefindings.

Source of Funding: None

MP39-03HIGH EXPRESSION OF HEME OXYGENASE-1 IS ASSOCIATEDWITH TUMOR PROGRESSION AND POOR CLINICAL OUTCOMEIN NON-MUSCLE INVASIVE BLADDER CANCER PATIENTS

Tomohiro Matsuo*, Yasuyoshi Miyata, Akihiro Asai, Kensuke Mitsunari,Kojiro Ohba, Hideki Sakai, Nagasaki, Japan

INTRODUCTION AND OBJECTIVES: Malignant aggressive-ness of bladder cancer is known to be regulated by cancer cell pro-liferation, angiogenesis, and lymphangiogenesis. These pathologicalprocesses were also controlled by various molecules including