Histone deacetylase inhibitor trichostatin A resensitizes gemcitabine resistant urothelial carcinoma cells via suppression of TG-interacting factor.

Gemcitabine and cisplatin (GC) has been widely used for advanced and metastatic urothelial carcinoma (UC). However, resistance to this remedy has been noticed. We have demonstrated that increase of TG-interacting factor (TGIF) in specimens is associated with worse prognosis of upper tract UC (UTUC) patients. The roles of TGIF in the gemcitabine resistance of UC were explored. Specimens of 23 locally advanced/advanced stage UTUC patients who received GC systemic chemotherapy after radical nephroureterectomy were collected to evaluate the alterations of TGIF in the resistance to the remedy by using immunohistochemistry. In vitro characterizations of mechanisms mediating TGIF in gemcitabine resistance were conducted by analyzing NTUB1 cells and their gemcitabine-resistant subline, NGR cells. Our results show that increased TGIF is significantly associated with chemo-resistance, poor progression-free survival, and higher cancer-related deaths of UTUC patients. Higher increases of TGIF, p-AKT(Ser473) and invasive ability were demonstrated in NGR cells. Overexpression of TGIF in NTUB1 cells upregulated p-AKT(Ser473) activation, enhanced migration ability, and attenuated cellular sensitivity to gemcitabine. Knockdown of TGIF in NGR cells downregulated p-AKT(Ser473) activation, declined migration ability, and enhanced cellular sensitivity to gemcitabine. In addition, histone deacetylases inhibitor trichostatin A (TSA) inhibited TGIF, p-AKT(Ser473) expression and migration ability. Synergistic effects of gemcitabine and TSA on NGR cells were also demonstrated. Collectively, TGIF contributes to the gemcitabine resistance of UC via AKT activation. Combined treatment with gemcitabine and TSA might be a promising therapeutic remedy to improve the gemcitabine resistance of UC.

Overexpression of TG-interacting factor is associated with worse prognosis in upper urinary tract urothelial carcinoma.

Prognostic outcome prediction would be useful for the treatment of patients with upper urinary tract urothelial carcinoma (UC). However, its prognostic biomarkers are not well established so far. According to the results of analysis of 168 human upper urinary tract UC specimens, overexpressed TG-interacting factor (TGIF) in nuclei of tumor tissues is significantly correlated with poor progression-free survival and higher cancer-related death. When both TGIF and p21 expression are altered, these patients had an even worse prognosis than those with one or no marker altered. Furthermore, to elucidate the role of TGIF in the progression of UC, overexpression of TGIF in RT4 or TSGH8301 cells was performed, and the results revealed that TGIF can significantly increase migration/invasion ability, matrix metalloproteinase expression, and invadopodia formation via the phosphatidylinositol 3-kinase-AKT pathway. In contrast, knockdown of TGIF with its specific short hairpin RNA inhibited the invasion ability of T24 cells. Besides, TGIF could inhibit p21(WAF/CIP1) expression, up-regulate cyclin D1 expression, and phosphorylate retinoblastoma to promote G1-S transition and cellular proliferation. In conclusion, we demonstrated that TGIF contributes to the progression of urothelial carcinoma via the phosphatidylinositol 3-kinase-AKT pathway. It may serve as an attractive therapeutic or prognostic target for selected patients with upper urinary tract UC.

Overexpression of annexin 1 in the development and differentiation of urothelial carcinoma.

This study investigates the expression of annexin 1 in urothelial carcinoma (UC) and its relation with clinicopathologic factors, and evaluates its potential clinical significance. Annexin 1 expression was analyzed by immunohistochemical staining with manual tissue microarrays and Western blot in UC. Immunohistochemical analysis of UC in tissue microarrays showed that annexin 1 protein was 76.5% (150/196) positive, which was markedly increased compared with that in the normal urothelium 20.8% (5/24) (p < 0.01). In addition, the positive expression rate of annexin 1 was higher in the high-grade UC (81.7%; 143/175) than in the low-grade UC (33.3%; 7/21). Western blot revealed that the expression of annexin 1 was low in low-grade UC, and markedly increased in high-grade UC. In conclusion, annexin 1 overexpression is observed in UC, which suggests it may be associated with tumorigenesis and its expression correlates with the differentiation of UC.

Influence of cytokine gene polymorphisms on prostate-specific antigen recurrence in prostate cancer after radical prostatectomy.

PURPOSE: Evidence is accumulating indicating that chronic inflammation plays an important role in prostate cancer. We investigated the potential prognostic roles of IL-6, IL-8 and IL-10 polymorphisms in clinical localized prostate cancer after radical prostatectomy. MATERIALS AND METHODS: A total of 116 clinically localized prostate cancer patients undergoing curative radical prostatectomy were included in this study. The IL-6, IL-8 and IL-10 polymorphisms were determined by the TaqMan real-time PCR method. Their prognostic significance on prostate-specific antigen (PSA) recurrence was assessed using Kaplan-Meier analysis and Cox regression model. RESULTS: The IL-6 polymorphism (rs2066992) T/G and G/G genotype cases were associated with a higher percentage of preoperative PSA levels of > or =10 ng/ml; higher risk of positive surgical margin, and higher risk of extraprostatic extension compared to the T/T genotype. The IL-10 polymorphism (rs1800871) A/A genotype was associated with a higher risk of PSA recurrence compared with the A/G + G/G genotypes and significantly poorer PSA-free survival (log-rank test, p = 0.019). After considering other covariates in a Cox proportional hazard model, the IL-10A/A genotype and high Gleason score (8-10) were still independent predictors of poor PSA-free survival. CONCLUSION: Our results suggest that the IL-10 polymorphism may be a prognostic factor for PSA recurrence after radical prostatectomy.

Overexpression of cyclooxygenase-2 in urothelial carcinoma in conjunction with tumor-associated-macrophage infiltration, hypoxia-inducible factor-1alpha expression, and tumor angiogenesis.

This study examines whether the expression of cyclooxgenase-2 (COX-2) in urothelial carcinoma (UC) is associated with macrophage infiltration, hypoxia-inducible factor-1alpha (HIF-1alpha) expression and angiogenesis. We investigated the expression of COX-2 associated with HIF-1alpha and performed double immunohistochemical analysis of 216 UCs for COX-2 expression and the correlation with tumor-associated-macrophage (TAM) density and microvessel density (MVD) in situ. A high expression of COX-2 was positively correlated with tumor invasiveness, histologic grade and HIF-1alpha expression in UC (p<0.0001, p=0.003, p<0.0001, respectively). Quantification of double staining of COX-2/CD34 and COX-2/CD68 showed that a higher MVD and TAM density was found in COX-2 high-expression than in COX-2 low-expression tumor fields (p<0.0001). Adjacent to the principal of COX-2 expression areas, MVD value and TAM density were significantly increased in HIF-1alpha high-expression specimens compared with HIF-1alpha low-expression ones (p<0.0001). Interestingly, our data revealed that high COX-2 expression (p=0.002), high HIF-1alpha expression (p<0.0001) and TAM density (p<0.0001) were all associated with high MVD value. Our results suggest that COX-2 may produce a cooperative effect in promoting tumor progression and may be involved in the process of angiogenesis through increasing TAM infiltration or HIF-1alpha regulation by hypoxia.

Experience with adjuvant chemotherapy for pseudomyxoma peritonei secondary to mucinous adenocarcinoma of the appendix with oxaliplatin/fluorouracil/leucovorin (FOLFOX4).

BACKGROUND: Pseudomyxoma peritonei (PMP) is a rare condition characterized by mucinous tumors, disseminated intra-peritoneal implants, and mucinous ascites. So far its diagnosis remains challenging to most clinicians. CASE PRESENTATION: A 55-year-old male patient had suffered from acute onset of abdominal pain and abdominal distension for one day prior to his admission. Physical examination revealed tenderness over the right lower quadrant of the abdomen without diffuse muscle guarding. A large amount of ascites was identified by abdominal computed tomography (CT) scan. Paracentesis showed the appearance of sticky mucinous ascites. He underwent laparotomy under the impression of pseudomyxoma peritonei. There was a lot of mucinous ascites, one appendiceal tumor and multiple peritoneal implants disseminated from the subphrenic space to the recto-vesicle pouch. Pseudomyxoma Peritonei caused by mucinous adenocarcinoma of appendiceal origin, was confirmed by histopathology. We performed an excision of the appendiceal tumor combined with copious irrigation and debridement. After the operation, he received 10 cycles of systemic chemotherapy with FOLFOX4 regimen, without specific morbidity. Follow-up of abdominal CT and colonoscopy at post-operative 17 months showed excellent response without evidence of local recurrence or distal metastasis. He made an uneventful recovery (up to the present) for 21 months after the operation. CONCLUSION: This case report emphasizes the possible new role of systemic chemotherapy in the treatment of patients with this rare clinical syndrome.

MDCT for differentiation of category T1 and T2 malignant lesions from benign gastric ulcers.

OBJECTIVE: The purpose of this study was to evaluate MDCT parameters for differentiating malignant (category T1 and T2) from benign gastric ulcers and to evaluate the performance characteristics of these predictors with optimal cutoff points determined in receiver operator characteristic analysis. SUBJECTS AND METHODS: The subjects were 26 patients with gastric cancer (11 with T1 lesions, 15 with T2 lesions) and 26 patients with benign gastric ulcer. MDCT and virtual gastroscopic findings were analyzed according to four qualitative criteria: ulcer shape, base, and margin and changes in adjacent folds. The quantitative criteria ulcer size, thickness of the gastric wall around an ulcer, thickness of the enhanced ulcer base, and enhancement around an ulcer were measured on multiplanar reconstruction images. We calculated the sensitivity and specificity of each quantitative criterion. Receiver operator characteristic analysis was used to identify cutoff points yielding optimal sensitivity and specificity for the diagnosis of gastric cancer. RESULTS: On virtual gastroscopy, ulcer shape and margin and gastric fold changes had sensitivities of 80.8%, 84.6%, and 90.9% and specificities of 76.9%, 73.1%, and 77.8%, respectively, in the diagnosis of gastric cancer. On multiplanar reconstruction images, thickness of the enhanced ulcer base and enhancement around the ulcer had sensitivities of 80.8% and 73.1% and specificities of 100% and 100%. CONCLUSION: MDCT combined with virtual gastroscopy and multiplanar reconstruction enhances the morphologic details of gastric ulcers and is a useful way to differentiate malignant (T1 and T2) and benign gastric ulcers.

Immunoexpression and prognostic role of hTERT and cyclin D1 in urothelial carcinoma.

The aim was to investigate the expression of human telomerase reverse transcriptase (hTERT) and cyclin D1 in correlation with clinicopathologic features of urothelial carcinoma (UC). Tissue microarrays (TMA) were constructed from paraffin-embedded specimens of 94 UC patients and immunohistochemical staining was used. High hTERT expression was found in 50 (53%) of the 94 tumors and was significantly associated with tumor invasiveness and tumor grade (P=0.008 and 0.0190, respectively). High cyclin D1 expression was found in 69 (73%) of the 94 tumors and was significantly associated with non-invasiveness and smaller tumor size, but there was no correlation with tumor grade. Kaplan-Meier analysis indicated that patients with low hTERT and high cyclin D1 levels had longer local recurrence-free survival (P=0.0482 and 0.0123, respectively). In addition, patients with high cyclin D1 levels had longer disease-free survival (P=0.0195). In conclusion, this study demonstrated that hTERT and cyclin D1 may be of recurrence predictive value for UC, thus providing clinicians with ancillary information when deciding on suitable therapeutic strategies in UC.

Cytologic diagnosis of primary effusion lymphoma in an HIV-negative patient.

Primary effusion lymphoma (PEL) is an unusual and rare type of non-Hodgkin's lymphoma characterized by lymphomatous effusion of pleural, pericardial or peritoneal cavities without lymphadenopathy or organomegaly. It is associated with human herpes virus-8 (HHV-8) and occurs most often in immunodeficient patients. We present a case of PEL in a 69-year-old male presenting with pleural effusion and ascites. Fluid aspiration showed a monomorphic population of atypical lymphoid cells, which were medium- to large-sized, with mono- or binucleated hyperchromatic nuclei and a small to moderate amount of basophilic cytoplasm containing cytoplasmic vesicles. Immunohistochemically, the lymphoid cells expressed CD138 and multiple myeloma oncogene 1, were positive for HHV-8, and were monoclonal for immunoglobulin heavy chain gene rearrangement. They were negative for Epstein-Barr virus by in situ hybridization. Unfortunately, the patient died during the first course of chemotherapy with cyclophosphamide, vincristine and prednisone.

Arsenic salt-induced DNA damage and expression of mutant p53 and COX-2 proteins in SV-40 immortalized human uroepithelial cells.

Arsenic is widely distributed in the environment, and is a proven toxic and carcinogenic agent that is associated with various human malignancies, including bladder cancer. However, the mechanisms of its carcinogenic action are still not well understood. In addition, over-expression of mutant p53 and cyclooxygenase-2 (COX-2) frequently occurs in a variety of human malignancies. It is therefore of interest to study the genotoxicity of arsenic salts on human uroepithelial cells and the expression of oncoproteins p53 and COX-2. In this study, the relative genotoxicity of sodium arsenite was evaluated in SV-40 immortalized human uroepithelial cells (SV-HUC-1) using the alkaline comet assay. The expression of mutant p53 and COX-2 was also evaluated by immunocytochemistry and western blotting. Our results revealed that sodium arsenite was able to induce DNA damage, and that its genotoxicity is correlated with its concentration. In addition, the expression of mutant p53 increased in parallel with comet scores, and the maximal expression of mutant p53 was observed at 4 microM arsenite. Similarly, sodium arsenite stimulated a concentration-dependent increase in COX-2 expression. In conclusion, this study demonstrated that sodium arsenite is genotoxic to uroepithelial cells in vitro, and that it will induce expression of mutant p53 and COX-2 proteins, indicating a possible key event in carcinogenesis. This study provides us with knowledge of the relationship between p53 and COX-2 over-expression in arsenite-treated urothelial cells and suggests a potential therapeutic role of COX-2 inhibitors in human urothelial malignancies.

Arsenic salts induced autophagic cell death and hypermethylation of DAPK promoter in SV-40 immortalized human uroepithelial cells.

Arsenic is a well-known toxic and carcinogenic agent, and associated with various human malignancies, including skin, lung and bladder cancers. Paradoxically, arsenic trioxide has been used successfully in the treatment of patients with acute promyelocytic leukemia. In addition, arsenic could induce cell apoptosis or autophagy in malignant cells. However, the underlying mechanism of arsenic-induced carcinogenesis is still unclear. In this study, we demonstrated an increase of autophagosomes was produced in arsenic-treated SV-HUC-1 cells by using electron microscopy. In addition, increase of Beclin-1, an important regulator for the formation of autophagosome, protein expression in a dose-dependent manner was also found. By using methylation specific PCR, we revealed hypermethylation of CpG sites in the promoter region with decreased DAPK protein expression in arsenic-treated SV-HUC-1 cells. As epigenetic silencing of tumor suppressor genes by promoter hypermethylation has been found in a variety of malignancies including bladder cancer, our results provide new insights for the understanding of the mechanism of arsenic-induced carcinogenesis in urothelial cells.

Expression of STAT3 and Bcl-6 oncoprotein in sodium arsenite-treated SV-40 immortalized human uroepithelial cells.

Arsenic is widely distributed in the environment, and it is a proven toxic and carcinogenic agent. On the southwest coast of Taiwan, an endemic occurrence of chronic arsenical poisoning due to a high concentration of arsenic in artesian-well water has been reported. However, the mechanisms of its carcinogenic action are still unclear. The Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway is an essential cascade for mediating normal functions of different cytokines in the development of the hematopoietic and immune systems. In this study, the substantial morphological changes observed in SV-40 immortalized human uroepithelial cells (SV-HUC-1) after treatment of various concentrations of arsenite were examined, and the expression of Bcl-6, Jak-2 and p-STAT3 (Tyr 705) were evaluated by immunocytochemistry and Western blotting. Our results showed that the expression of Bcl-6 increased dose-dependently in arsenite-treated urothelial cells. Sodium arsenite treatment reduced Jak-2 protein expression in a dose-dependent manner. However, treatment of SV-HUC-1 cells with arsenite at concentration ranges from 2 and 4microM for 48h dramatically increased p-STAT3 (Tyr 705), but the levels decreased at 8-40microM of arsenite. Our data suggest that arsenic-mediated inactivation of the JAK-STAT signaling pathway might be caused by Bcl-6 interaction with JAK tyrosine kinase or STAT. In conclusion, our findings indicate that arsenic inhibits JAK tyrosine kinase protein expression and suggest the interference in the JAK-STAT pathway might be through Bcl-6 playing an important role in arsenic-associated carcinogenesis.

p-Phenylenediamine induced DNA damage in SV-40 immortalized human uroepithelial cells and expression of mutant p53 and COX-2 proteins.

p-Phenylenediamine (p-PD) is the main aromatic amine used in the formulation of hair dyes. Some epidemiologic studies have suggested that the use of p-PD-based hair dyes might be related to increased risk of human malignant tumors including bladder cancer and hematopoietic cancers. However, the toxicity and genotoxicity of p-PD on urothelial cells has not been reported yet. Therefore, we investigated the genotoxicity of p-PD on human urothelial cells and study its association with the expression of oncoproteins p53 and cyclooxygenase-2 (COX-2). Our results revealed that p-PD was able to induce DNA damage determined by Comet assay. In addition, our immunocytochemical and Western blotting results showed that p-PD induced overexpression of mutant p53 and COX-2 in a dose-dependent manner. The relationship between mutant p53 and COX-2 expression shows strong correlation. Furthermore, the accumulation of mutant p53 was linearly correlated with Comet scores. These results suggest that p-PD can induce DNA damage and accumulation of mutant p53 and COX-2 proteins; this may be one of the possible mechanisms that cause genotoxic carcinogenesis in the urothelial cells.

Gastric cancer: preoperative local staging with 3D multi-detector row CT--correlation with surgical and histopathologic results.

PURPOSE: To prospectively evaluate accuracy of multi-detector row computed tomographic (CT) images for preoperative staging of gastric cancer by using surgical and histopathologic results as reference standards. MATERIALS AND METHODS: This study had institutional review board approval; informed consent was obtained from all patients. Multi-detector row CT included acquisition of virtual gastroscopy images after air distention and contrast material-enhanced dynamic transverse and multiplanar reformation (MPR) images after water distention. Fifty-five consecutive patients with gastric cancer (38 men, 17 women; age range, 37-84 years; mean age, 63 years) underwent preoperative CT. All received 6 g of gas-producing crystals before unenhanced CT scanning for gastric distention and virtual gastroscopy. Patients drank 800-1000 mL of tap water to establish a background for dynamic contrast-enhanced CT scans. Images were obtained in late arterial, portal venous, and delayed phases with start delays of 40, 70, and 150 seconds, respectively. All patients underwent surgery. CT findings were compared with surgical and histopathologic results. Differences in accuracy of transverse and MPR images for T and N staging were assessed with the McNemar exact test. Statistical significance was inferred at P < .05. RESULTS: Detection rates of primary tumors with transverse images, MPRs, and combinations of MPR and virtual gastroscopy images were 91% (50 of 55), 96% (53 of 55), and 98% (54 of 55), respectively. Overall accuracy in assessment of tumor invasion of the gastric wall (T stage) was significantly better with MPR images (89% [49 of 55]) than with transverse images (73% [40 of 55]) (P < .01). Overall accuracy for lymph node (N) staging was 78% (43 of 55) with MPR images and 71% (39 of 55) with transverse images. This difference was not significant (P = .103). CONCLUSION: Multi-detector row CT with combined water and air distention can improve the accuracy of preoperative staging of gastric cancer. MPRs yield significantly better overall accuracy than transverse images for tumor staging but not for lymph node staging.

The expression of CD66a and possible roles in colorectal adenoma and adenocarcinoma.

BACKGROUND: The aim of our study was to analyze the expression and possible role of CD66a in colorectal adenoma and adenocarcinoma and the relationship between its expression and pre-operation serum carcinoembryonic antigen (CEA) level and tumor stage in patients with colorectal adenocarcinomas. METHODS: Paraffin-embedded sections from 184 patients including 42 colorectal adenomas with low-grade dysplasia, 43 adenomas with high-grade dysplasia, and 99 adenocarcinomas were collected for this study. Immunohistochemical analysis was performed, and the expression and the location of CD66a were evaluated and were correlated with beta-catenin nuclear expression. RESULTS: The expression of CD66a was found not only in the apical membrane of neoplastic glands but also in secretion within the lumen of the neoplastic glands including adenomas and adenocarcinomas. Expressions of secreted CD66a were of higher level in adenocarcinoma than in adenoma with high-grade dysplasia and adenoma with low-grade dysplasia (p < 0.0001). High expression of secreted CD66a was also associated with tumor stage, invasion, and pre-operation serum CEA level of patients with colorectal adenocarcinoma. CONCLUSIONS: This study implied that CD66a can function both as an epithelial cell adhesion protein or alternatively as secreted CD66a. In addition, a high expression of CD66a was significantly correlated with tumor invasion, stage, and pre-operation serum CEA level.

Primary pulmonary synovial sarcoma: a case report.

We report a rare case of primary synovial sarcoma of the lung. A57-year-old man had a well-defined tumor in the right middle lobe seen on chest computed tomography, and underwent lobectomy. Grossly, the nonencapsulated tumor measured 4.5 cm in greatest diameter, with a solid and tan-white cut surface. Histologically, the tumor was mainly composed of a dense proliferation of spindle cells. Immunohistochemical studies were focally positive for epithelial membrane antigen, and diffusely positive for CD99 and Bcl-2. Cytokeratin, S-100 protein, desmin, smooth muscle act in, and CD34 were absent. SYT-SSX1 gene fusion transcript was detected by reverse transcription-polymerase chain reaction, which is diagnostic of primary synovial sarcoma of the lung. We also review the literature with regard to the clinicopathologic, immunohistochemical, and molecular studies of primary pulmonary synovial sarcoma.

Clinical and radiographic presentations of pulmonary cryptococcosis in immunocompetent patients.

Pulmonary cryptococcosis is often noted in immunocompromized patients, especially in HIV-seropositive patients and post-transplant patients. Only a few case reports and small-scale studies on pulmonary cryptococcosis in immunocompetent patients have been published in the English literature. However, there are several areas of uncertainty in this group of patients including image presentations and management strategy. This retrospective study including 17 patients is designed to share our 7 y of experience in clarifying the characteristics of pulmonary cryptococcosis in immunocompetent patients, including initial symptoms, diagnostic criteria, pulmonary imaging, treatment and outcome in a tertiary teaching hospital in Taiwan.

Huge aggressive angiomyxoma: a case report and literature review.

Angiomyxoma occurs mostly in 30- to 40-year-old females and is described histologically as a mesenchymal tumor, composed of fibroblasts within a strong myxoid background. It occurs mainly in the female pelvis, vulva or perineum, and grows slowly. Treatment is surgical excision. Unfortunately, there is a relatively high rate of recurrence because the exact extent of the tumor is difficult for the surgeon to determine. We report a case of aggressive angiomyxoma combined with uterine myoma, and discuss the characteristics of its images. Surgical excision of the tumor was performed, and adjuvant treatment was given for local recurrence.

Mature cystic teratoma in the anterior mediastinum containing a carcinoid.

Mature teratomas are common neoplasms in the anterior mediastinum. However, a primary carcinoid tumor occurring in a mature teratoma is extremely rare. To our knowledge, there are only 2 published articles in the literature to date reporting a mature cystic teratoma of the mediastinum containing a carcinoid. We herein report another case of primary carcinoid tumor arising from a mature cystic teratoma in the anterior mediastinum.