Takuji Yasuda

Pathological significance and prognostic implications of heme oxygenase 1 expression in non‑muscle‑invasive bladder cancer: Correlation with cell proliferation, angiogenesis, lymphangiogenesis and expression of VEGFs and COX‑2

Heme oxygenase 1 (HO-1) is a stress-response protein and its expression is associated with malignant potential and poor prognosis in several types of cancer. The present study investigated the association between HO-1 expression levels and the pathological features, clinical outcomes and other associated factors in patients with non-muscle-invasive bladder cancer (NMIBC). HO-1 expression was evaluated using immunohistochemistry in 147 formalin-fixed tissue specimens. The proliferation index, microvessel density, lymph vessel density and expression of cyclooxygenase (COX)-2 and vascular endothelial growth factor (VEGF)-A, -C, and -D were also investigated. Correlations among variables were analyzed by multivariate analysis. Survival was assessed using Kaplan-Meier survival curves and multivariate statistics. HO-1 expression levels in high-grade and pT1 tumors were significantly higher compared with low-grade and pTa tumors, and were correlated with the proliferation index (P<0.001), lymph vessel density (P=0.021) and COX-2 expression levels (P=0.003). The proliferation index and COX-2 expression levels were also identified as independent contributing factors in multivariate models. Kaplan-Meier survival curves associated HO-1 expression with a poor prognosis in metastasis-free (P=0.047) and cause-specific survival (P=0.017), but not with urinary tract recurrence (P=0.231). Furthermore, HO-1 expression was identified by multivariate analysis to be a significant predictor for cause-specific survival (hazard ratio, 4.08; 95% confidence interval, 1.06-15.66; P=0.004). HO-1 has an important role in the malignant aggressiveness of NMIBC and its expression is associated with cause-specific survival. HO-1-associated activities are regulated by cancer cell proliferation, lymphangiogenesis and COX-2. The results suggest that HO-1 may be a potential therapeutic target and a useful predictive prognostic factor in patients with NMIBC.

Stromal expression of Fer suppresses tumor progression in renal cell carcinoma and is a predictor of survival

Fps/Fes related (Fer) is a non-receptor tyrosine kinase that is expressed in fibroblasts, immune cells and endothelial cells. Fer serves an important pathological role in cell survival, angiogenesis and the immune system. However, the pathological role of Fer expression in the stromal cells surrounding renal cell carcinoma (RCC) has not been previously investigated. In the present study, immunohistochemical analysis of Fer was performed using the formalin-fixed tissue samples of 152 patients with RCC. The proliferative and apoptotic indices were used to represent the percentage of proliferation marker protein Ki-67- and cleaved caspase-3-positive cells, respectively. The microvessel density was defined as the number of cluster of differentiation (CD) 31-positively stained vessels/mm2. In addition, CD57+ and CD68+ cells were counted using semi-quantification of natural killer (NK) cells and macrophages. Fer expression in stromal cells was negatively associated with Fuhrman grade, pathological tumor stage and metastasis (P<0.001). Fer expression in stromal cells was negatively associated with CD68+ macrophage density, whereas it was positively associated with CD57+ NK cell density. Kaplan-Meier estimators indicated that decreased stromal Fer expression was a predictive marker of decreased cause-specific survival rate (P<0.001). Furthermore, low expression of Fer was identified as being an independent marker of decreased cause-specific survival using multivariate analysis (hazard ratio, 7.4; 95% confidence interval, 1.7-33.0; P<0.001). The results of the present study suggested that low Fer expression in stromal cells is associated with increased malignant aggressiveness and decreased survival in patients with RCC. CD57+ NK cell and CD68+ macrophage regulation in cancer-stromal tissue is considered to affect RCC pathology.

High Consumption of Green Tea Suppresses Urinary Tract Recurrence of Urothelial CancerviaDown-regulation of Human Antigen-R Expression in Never Smokers

Background/Aim: Smoking is a risk factor for carcinogenesis and progression of urothelial cancer (UC). Green tea polyphenol inhibits these malignant behaviors and suppresses human antigen R (HuR) expression, which is associated with malignant aggressiveness. This study aimed to clarify the anti-cancer effects of green tea based on the smoking status of UC patients. Patients and Methods: Three hundred and sixty (260 with bladder cancer, BC and 100 with upper tract UC) patients were divided into three groups based on consumption of green tea: low (<1 cup/day, n=119), middle (1-4 cup/day, n=160), and high (>5 cup/day, n=81). HuR immunoreactivity was evaluated immunohistochemically in formalin-fixed specimens. Results: In never smokers, multivariate analysis showed that the frequency of green tea consumption was a significant predictor (middle: hazard ratio, HR, 0.36, p=0.002; high: HR, 0.20, p=0.003) of urinary tract recurrence. A high consumption of green tea was associated with low rates of urinary tract recurrence and up-grading in UC patients. In BC, high consumption was associated with a lower risk of up-grading (p=0.011) and up-staging (p=0.041) in recurrent cancer. HuR expression in the high-consumption group was lower (p=0.019) than that in other groups. These significant findings were not detected in ever smokers. Conclusion: High consumption of green tea suppressed urinary tract recurrence and the risks of up-grading and up-staging by recurrence in never smokers. Our results suggested that HuR expression played important roles in such mechanisms.

MP65-01 4N1K-PEPTIDE DERIVED FROM THROMBOSPONIDN-2 IS ASSOCIATED WITH MALIGNANT AGGRESSIVENESS AND PROGNOSIS IN BLADDER CANCER

RESULTS: Median follow-up of the series was 131.8 months (120-194). During the surveillance period, 55 patients developed BCR (33%) and 6 metastatic recurrence (3.6%). Overall, three patients died of PCa (1.8%), and 22 due to other causes (13%). Expression levels of 15800 genes differed between clinically localized and locally advancedmetastatic PCa groups (FDR<0.1). A total of 35 differently expressed genes with a fold change value > 5 and 10 genes previously described in literature were selected to be validated in an additional set of samples. Two genes were found to be independent prognostic factors of BCR (MSMB, HR1⁄41.75, p1⁄40.04) (FOS, HR1⁄41.73, p1⁄40.05) and two genes were found to be predictors of metastatic recurrence (KRT17, HR1⁄47.17, p1⁄40.03) (SERPINE1, HR1⁄45.54, p1⁄40.06). CONCLUSIONS: Gene expression levels in PCa tissue can be useful for distinguishing patients with clinically localized disease who will develop BCR or metastatic recurrence after radical prostatectomy. These gene expression biomarkers could have potential clinical utility for identifying the subset of patients that would benefit from closer surveillance and adjuvant therapy.

MP54-07 HEPATOCYTE GROWTH FACTOR RECEPTOR/C-MET IS ASSOCIATED WITH MALIGNANT AGGRESSIVENESS VIA REGULATION OF CYCLOOXYGENASE-2, HEMEOXYGENASE-1, AND PROGRAMMED DEATH LIGAND 1 IN PATIENTS WITH BLADDER CANCER

(IHC) on tissue microarrays were used to confirm tissue sampling and the gene expression analysis. RESULTS: Unsupervised consensus clustering yielded four distinct consensus clusters (CC) or subtypes: CC1-Basal, CC2Luminal, CC3-Immune and CC4-Scar-like. The CC1-Basal and CC2Luminal subtypes expressed genes consistent with a basal-like (KRT5/ 6, KRT14) and a luminal-like (GATA3, PPARG) phenotype, respectively. The CC3-Immune subtype had the highest immune activity, including T-cell infiltration and checkpoint molecules (CTLA4, CD80). Finally, the CC4-Scar-like profile was consistent with non-neoplastic scar samples, expressing genes associated with wound healing / scarring (MYH11, SHH, CNN1). Approximately the same numbers of pre-NAC basaland luminal-like samples remained static (CC1 or CC2) with respect to subtype or were highly immune-infiltrated (CC3) in the post-NAC setting. In the post-NAC setting, luminal-like pre-NAC samples were more likely adopt a scar-like character (CC4). CONCLUSIONS: This study expands our knowledge of cisplatin-resistant MIBC by suggesting molecular subtypes to understand the biology of these tumors. Although these molecular subtypes imply consequences for adjuvant treatments, this ultimately needs to be tested in clinical trials.

Pathological significance and prognostic roles of densities of CD57+ cells, CD68+ cells, and mast cells, and their ratios in clear cell renal cell carcinoma

The immune system is closely associated with malignant behavior in renal cell carcinoma (RCC). Therefore, understanding the pathological roles of immune cells in tumor stroma is essential to discuss the pathological characteristics of RCC. In this study, the clinical significance of densities of CD57+ cells, CD68+ cells, and mast cells, and their ratios were investigated in patients with clear cell RCC. The densities of CD57+, CD68+, and mast cells were evaluated by immunohistochemical techniques in 179 patients. Proliferation index, apoptotic index, and microvessel density were evaluated by using anti-Ki-67, anti-cleaved caspase-3, and anti-CD31 antibodies, respectively. The density of CD57+ cell was negatively correlated with grade, pT stage, and metastasis, although densities of CD68+ cell and mast cell were positively correlated. Ratios of CD68+ cell/CD57+ cell and mast cell/CD57+ cell were significantly correlated with grade, pT stage, and metastasis. Survival analyses showed that the CD68+ cell/CD57+ cell ratio was a significant predictor for cause-specific survival by multivariate analyses (hazard ratio = 1.41, 95% confidence interval = 1.03-1.93, P = .031) and was significantly correlated with proliferation index, apoptotic index, and microvessel density (r = .47, P <. 001; r = -.31, P < .001; and r = .40, P < .001, respectively). In conclusion, CD57+ cells, CD68+ cells, and mast cells played important roles in malignancy in clear cell RCC. The CD68+ cell/CD57+ cell ratio was strongly correlated with pathological features and prognosis in these patients because this ratio reflected the status of cancer cell proliferation, apoptosis, and angiogenesis.

Efficacy and safety of sunitinib alternate day regimen in patients with metastatic renal cell carcinoma in Japan: Comparison with standard 4/2 schedule

Sunitinib is a standard agent for metastatic renal cell carcinoma (mRCC). The standard schedule, 4 weeks‐on followed by 2 weeks‐off (4/2 schedule), often does not maintain an adequate dosage because of the severe adverse events (AEs). We compared the efficacy and safety of an alternative every other day (q.a.d.) dosing with that of the 4/2 schedule in mRCC patients.

Neoadjuvant hormonal therapy for low-risk prostate cancer induces biochemical recurrence after radical prostatectomy via increased lymphangiogenesis-related parameters

The effects of neoadjuvant hormonal therapy (NHT) on pathological features and lymphangiogenesis in patients with prostate cancer (PCa) for each pre‐operative risk classification are unclear.

MP88-02 EXPRESSION OF CLASS III BETA-TUBULIN PREDICTS PROGNOSIS IN CISPLATIN-RESISTANT BLADDER CANCER PATIENTS RECEIVING PACLITAXEL-BASED SECOND-LINE CHEMOTHERAPY

INTRODUCTION AND OBJECTIVES: Prior experimental studies have shown that the enzyme g-glutamyltransferase (g-GT) is overexpressed during carcinogenetic transformation of urothelial cells. This analysis aimed to elucidate the prognostic value of serum g-GT, glutamat-pyruvat-transaminase (GPT) and glutamat-oxalattransaminase (GOT) levels in patients with invasive bladder cancer (BC). METHODS: Preoperative serum g-GT, GPT and GOT concentrations were assessed in 324 patients treated with RC for clinically non-metastatic BC between 2002 and 2013. Laboratory values were obtained 1-3 days prior to RC. Uniand multivariable analyses were carried out to evaluate clinicopathologic features and survival after RC. The median follow-up was 36 months (IQR: 10-55). RESULTS: Elevated preoperative g-GT, GPT and GOT levels were diagnosed in 77 (23.8%), 20 (6.2%) and 18 (5.5%) patients, respectively. Elevated g-GT levels were significantly associated with advanced tumor stage ( pT3a; p1⁄40.001), lymph-node tumor involvement (p<0.001), positive surgical margins (p1⁄40.018), lymphovascular invasion (p1⁄40.024), muscle-invasive disease at primary diagnosis (p1⁄40.033), increased tumor size (continuously coded; p1⁄40.035), receipt of neoadjuvant chemotherapy (p1⁄40.006), higher Eastern Cooperative Performance Status (p1⁄40.001), hydronephrosis at RC (p1⁄40.049), increased preoperative serum C-reactive protein levels (p<0.001) and elevated serum GPT and GOT levels (both p<0.001). Patients with elevated g-GT concentration exhibited inferior 3-year disease-free (52.5% vs. 63.2%; p1⁄40.19), cancer-specific (71.1% vs. 80.9%; p1⁄40.042) and overall survival rates (49.2% vs. 69.6%; p1⁄40.005) compared to patients with normal levels. On multivariable analysis, higher ECOG performance status, hydronephrosis at RC (both p1⁄40.010), positive surgical margin status (p1⁄40.014), lymph node positive disease (p1⁄40.030), advanced tumor stage (p1⁄40.032), and an elevated g-GT concentration (p1⁄40.043) were independent predictors of all-cause mortality. CONCLUSIONS: Elevated preoperative g-GT levels are associated with a significantly increased risk for locally advanced bladder cancer and mortality. These data suggest that g-GT levels may be a useful prognostic marker for patients after RC for BC. Source of Funding: None