Analiza ekspresije transformišućeg faktora rasta beta i ćelijskih receptora smrti u urotelnom karcinomu mokraćne bešike i njihov prognostički značaj

Bladder cancer is the most frequent malignant tumor of the urinary tract. It represents a significant burden to the healthcare system due to necessity of lifelong cystoscopic controls, and management of recurrent disease. More than 90% of bladder cancers are urothelial carcinomas. Dysregulation of transforming growth factor beta (TGF- β) signaling pathways plays important role in tumor development and progression. Activation of TGF-β and cell death receptors involves common pathways in the activation of caspases and induction of apoptosis. The significance of immunohistochemical expression of TGF-β and death receptors in urothelial bladder cancer has not been fully elucidated. This study comprised 647 urothelial bladder cancers obtained by transurethral resection that were immunohistochemically analyzed to the expression of TGF-β1 and death receptors DR4, DR5, and FAS. The expression of Smad4, a protein with the central role in TGF-β canonical pathway signal transmission, and EZH2, an epigenetic regulator involved in TGF-β signalization, was also analyzed. Association assessment between the markers’ expression and clinicopathological parameters and follow-up data was performed. High TGF-β1 expression in urothelial cancer was significantly associated with high tumor grade and advanced stage, while the reduction/loss of death receptors’ expression correlated to muscle-invasive disease. High Smad4 expression was associated with tumor recurrence. Cancer-specific mortality was directly linked to high TGF-β1 and EZH2 expression, and inversely to DR4, FAS, and Smad4 expression. High TGF-β1 was associated with shorter overall survival of the patients, while high expression of DR4 and FAS in tumor cells correlated with less aggressive phenotype of the tumor, and longer overall survival. The present study identified the expression of TGF-β1, FAS, and EZH2 as independent prognostic factors in urothelial bladder cancer, where high TGF-β1 and EZH2 expression indicates shorter survival and poor prognosis, while high FAS receptor expression imparts protective aspect and is linked to longer overall survival. Further insight into knowledge about complex roles and significance of TGF-β and death receptors, as well as the analysis of their expression in urothelial bladder cancer, may have important implications for prognostic stratification of the patients and in deciding about their clinical management

Stromal reaction and prognosis in acinic cell carcinoma of the salivary gland

Introduction. Primary acinic cell carcinoma (ACC) is an uncommon malignant neoplasm of the salivary gland (SG), which usually presents as slow growing tumor. Case report. We reported a 69-year-old woman with tumor in the right parotid gland with a 5-year progress. Biopsy sections revealed a hybrid form of ACC with a low- and high-grade component and prominent lymphoid tissue in tumor stroma. Immunohistochemistry was performed to define the molecular profile of this unusual ACC, with special interest for stromal influence on to the proliferative activity of ACC with dedifferentiation. We detected that the level and the type of stromal lymphoid reaction (particularly CD8+/CD4+ ratio) had a significant influence on to Ki-67 index in the high-grade component of ACC, as well as the involvement of the CXCR4 signaling axis in the stromal reaction influence. Conclusion. We suggest that tumor stroma may be a source of potential new tumor biomarkers which can determine the aggressivity of this tumor.[Projekat Ministarstva nauke Republike Srbije, br. 175092

Pro- and Antiapoptotic Markers in Upper Tract Urothelial Carcinoma Associated with Balkan Endemic Nephropathy

The role of aristolochic acid in the etiology of Balkan endemic nephropathy (BEN) and associated upper-tract urothelial carcinoma (UTUC) has been recently confirmed. The aim of this study was to determine apoptosis-related marker(s) specific for BEN-associated UTUC. Present investigation included 105 patients with UTUC, 44 from BEN region and 61 control tumors. Altered expression of Survivin was more often present in BEN UTUC with high grade and solid growth (P < 0.005; P < 0.05) than in control tumors. Significantly lower expression of proapoptotic marker Bax was found in BEN tumors with high grade, high stage, necrosis, and without metaplastic change (P < 0.05; 0.05; 0.05; 0.05) compared to control tumors with the same features. Group (BEN-related/control), stage, growth pattern, and caspase 3 activity were significantly associated with the expression of Bax (P = 0.002, 0.034, 0.047, 0.028, resp.,). This investigation identifies Bax as specific marker of BEN-associated UTUC. Decrease of pro-apoptotic protein Bax together with alteration of Survivin may be indicative for specific disturbances of intrinsic apoptotic pathway in UTUC arising in endemic areas

The Encapsulation of Lycopene in Nanoliposomes Enhances Its Protective Potential in Methotrexate-Induced Kidney Injury Model

Methotrexate is an antimetabolic drug with a myriad of serious side effects including nephrotoxicity, which presumably occurs due to oxidative tissue damage. Here, we evaluated the potential protective effect of lycopene, a potent antioxidant carotenoid, given in two different pharmaceutical forms in methotrexate-induced kidney damage in rats. Serum biochemical (urea and creatinine) and tissue oxidative damage markers and histopathological kidney changes were evaluated after systemic administration of both lycopene dissolved in corn oil and lycopene encapsulated in nanoliposomes. Similar to previous studies, single dose of methotrexate induced severe functional and morphological alterations of kidneys with cell desquamation, tubular vacuolation, and focal necrosis, which were followed by serum urea and creatinine increase and disturbances of tissue antioxidant status. Application of both forms of lycopene concomitantly with methotrexate ameliorated changes in serum urea and creatinine and oxidative damage markers and markedly reversed structural changes of kidney tissue. Moreover, animals that received lycopene in nanoliposome-encapsulated form showed higher degree of recovery than those treated with free lycopene form. The findings of this study indicate that treatment with nanoliposome-encapsulated lycopene comparing to lycopene in standard vehicle has an advantage as it more efficiently reduces methotrexate-induced kidney dysfunction

The association between NOTCH3 expression and the clinical outcome in the urothelial bladder cancer patients

Disrupted NOTCH activity is a driving event in urothelial bladder cancer (UBC). After activation by hypoxia, the NOTCH3 receptor participates in tumor cell proliferation, acquisition of the epithelial-mesenchymal transition phenotype, and angiogenesis. The aim was to analyze the association of NOTCH3 expression with histopathological and clinical parameters, and to determine its predictive impact on the clinical outcome in UBC patients. The present research included 614 UBC samples incorporated in paraffin tissue microarrays, evaluated by immunohistochemistry for NOTCH3 expression. The accrual period was four years, while the follow-up period was two years. The membranous expression was semi-quantified (0-3), and the mean degree was 1.81±0.94. Criteria for semi-quantification the NOTCH3 expression were the intensity of the staining and the percentage of positive cells. The samples with negative (0) and weak (1) NOTCH3 immunohistochemical (IHC) score were considered negative, while the samples that showed moderate (2) and strong (3) expression were considered positive. Higher degree of positivity was associated with higher risk of cancer-specific mortality (p<0.001). Independent predictors for cancer-specific mortality were NOTCH3 expression and high stage (p<0.001). NOTCH3 expression was not a statistically significant predictor of recurrence-free survival (p=0.816). This study indicated that NOTCH3 is a predictor of poor outcome, suggesting that the NOTCH3 could be potentially reliable IHC marker for selecting the UBC patients that would require more intensive follow-up, especially if they diagnosed in higher stage, with divergent differentiation in pathological report, and without recurrences which would lead them to more frequent medical assessments

Original Article Angiogenesis in upper tract urothelial carcinoma associated with Balkan endemic nephropathy

Abstract: Upper tract urothelial carcinoma (UTUC) associated with Balkan endemic nephropathy (BEN) is characterized by a number of aberrations in cell-cycle regulation and apoptosis. The aim of this study was to detect angiogenesis-related marker(s) specific for BEN UTUC, and to examine the influence of HIF 1α upon angiogenesis and apoptosis in UTUC. Present investigation included 110 patients with UTUC, 50 from BEN region and 60 control tumors. Altered expression of VEGFR1 was more often present in control UTUC than in BEN tumors (p&lt;0.005). It was associated with high grade, low and high stage, solid growth, and metaplastic change of control UTUC. Microvessel density assessed by CD31 (MVD CD31) was significantly higher in UTUC with lymphovascular invasion (p&lt;0.05), and in BEN tumors with papillary growth (p&lt;0.05). Discriminant analysis indicated that BEN and control tumors do not differ significantly in expression of angiogenesis related markers. The most important discriminant variable that determined control UTUC was expression of VEGFR1 (p=0.002). HIF 1α in UTUC significantly correlated with the low stage, papillary growth and expression of Bcl-2, Caspase-3 index, and MVD CD34 (p&lt;0.001; 0.0005; 0.01; 0.005; 0.01, respectively). HIF-1α may be helpful marker in evaluation of UTUC, especially when combined with angiogenesis and apoptosis

Analiza ekspresije transformišućeg faktora rasta beta i ćelijskih receptora smrti u urotelnom karcinomu mokraćne bešike i njihov prognostički značaj

Bladder cancer is the most frequent malignant tumor of the urinary tract. It represents a significant burden to the healthcare system due to necessity of lifelong cystoscopic controls, and management of recurrent disease. More than 90% of bladder cancers are urothelial carcinomas. Dysregulation of transforming growth factor beta (TGF- β) signaling pathways plays important role in tumor development and progression. Activation of TGF-β and cell death receptors involves common pathways in the activation of caspases and induction of apoptosis. The significance of immunohistochemical expression of TGF-β and death receptors in urothelial bladder cancer has not been fully elucidated. This study comprised 647 urothelial bladder cancers obtained by transurethral resection that were immunohistochemically analyzed to the expression of TGF-β1 and death receptors DR4, DR5, and FAS. The expression of Smad4, a protein with the central role in TGF-β canonical pathway signal transmission, and EZH2, an epigenetic regulator involved in TGF-β signalization, was also analyzed. Association assessment between the markers’ expression and clinicopathological parameters and follow-up data was performed. High TGF-β1 expression in urothelial cancer was significantly associated with high tumor grade and advanced stage, while the reduction/loss of death receptors’ expression correlated to muscle-invasive disease. High Smad4 expression was associated with tumor recurrence. Cancer-specific mortality was directly linked to high TGF-β1 and EZH2 expression, and inversely to DR4, FAS, and Smad4 expression. High TGF-β1 was associated with shorter overall survival of the patients, while high expression of DR4 and FAS in tumor cells correlated with less aggressive phenotype of the tumor, and longer overall survival. The present study identified the expression of TGF-β1, FAS, and EZH2 as independent prognostic factors in urothelial bladder cancer, where high TGF-β1 and EZH2 expression indicates shorter survival and poor prognosis, while high FAS receptor expression imparts protective aspect and is linked to longer overall survival. Further insight into knowledge about complex roles and significance of TGF-β and death receptors, as well as the analysis of their expression in urothelial bladder cancer, may have important implications for prognostic stratification of the patients and in deciding about their clinical management

TP53 GENE MUTATIONS – FROM GUARDIAN OF THE GENOME TO ONCOGENE

TP53 tumor suppressor gene mutations are the most frequent genetic alterations in human cancer affecting a specific gene. The occurrence of TP53 mutations is considerably influenced by cancer-initiating events, such as DNA damage, the aftermath of which is the promotion of cancer development through the loss of anti-proliferative activities, including apoptosis and cellular senescence. Over 27.000 TP53 gene mutations have been discovered and found in more than 50% of human cancers. The most frequent alterations are the point mutations with a single base substitution in gene segment encoding for DNA-binding domaine of p53 molecule, leading to the production of mutant protein that differs from the wild-type protein by one amino acid (missense mutations) usually causing the change in tertiary structure of gene product, thus preventing p53 to bind to DNA and activate transcription of target genes. The result of the mutations may also be the proteins with new, abnormal functions, and the ability to modulate expression of genes responsible for neoangiogenesis, resistance to chemotherapeutics and prevention of tumor initiation and promotion. In such circumstances, not only the mutant TP53 loses its tumor suppressive function, but acquires oncogenic potential and becomes an active participant in the neoplastic transformation of the cell.Vast heterogeneity of mutations and methodological approaches in p53 status assessment represent the main difficulties in rapid and effective integration of basic p53 research into clinical practice

Malakoplakia mimics urinary bladder cancer: A case report

Introduction. Malakoplakia is an unusual and very rare chronic inflammatory disease. In bladder especially it can mimic malignancy and lead to serious misdiagnosis. Case report. We presented a case of a middle-aged woman with persistent macrohematuria and cystoscopically polypoid bladder mass that resembled a neoplastic process. The final diagnosis was based on cystoscopic biopsy and microscopic findings of acidophilic, foamy histiocytes with the presence of Michaelis-Gutmann inclusions which are characteristic for diagnosis of malakoplakia. Immunohistochemistry confirmed diagnosis by demonstrating CD68-positive macrophages. Conclusion. Urinary bladder malakoplakia should be considered in patients with persistent urinary tract infections and tumor mass at cystoscopy. Early identification with prompt antibiotic treatment can be helpful in avoiding unnecessary surgical interventions and in preventing development of possible complications. [Projekat Ministarstva nauke Republike Srbije, br. 175092

Synchronous mantle cell lymphoma and prostate adenocarcinoma-is it just a coincidence?

Introduction. Synchronous occurrence of lymphomas and other cancers, mostly carcinomas are well established. The most of cases describe chronic lymphocytic leukemia as the leading lymphoproliferative disease with the tendency towards secondary malignancies development. Mantle cell lymphoma (MCL) has been described in only 2 cases to co-occur with prostate adenocarcinoma (PAC). There are scarce data about the connection between MCL and urology cancers. We presented the first case of synchronous occurrence of MCL and PAC in the same patient in Serbia. Case report. A 64-year-old male initially presented with fatigue, splenomegaly, and bicytopenia. The bone marrow biopsy specimen revealed extensive infiltration with MCL. During lymphoma staging procedure prostate enlargement (57 mm) was accidentally found by multislice- computed tomography (MSCT). The serum prostate specific antigen (PSA) was elevated (52 ng/mL; normal values ≤ 4 ng/mL). Transrectal ultrasound biopsy revealed PAC. High Gleason score determined high-risk locally advanced PAC. The patient underwent treatment with chemotherapy and hormone therapy due to the existence of double malignancies. Cyclophosphamide, doxorubicin, vincristine, prednisone (CHOP) was applied for MCL, and luteinizing hormonereleasing hormone (LHRH) agonist, triptorelin, for PAC. Partial response was obtained for MCL, and stable disease for PAC. In a 1.5-year observation period the patient was still disease progression free for both of malignancies. Conclusion. This case points aut that elderly males are in need for careful observation during the staging procedure for lymphoma. The literature data suggest that MCL patients are in increased risk for urologic malignancies development. However, the etiologic connection between these two entities, except male gender and older age, remains unclear