22 research outputs found
Comparative study of the expression of RHO kinases in urinary bladder cancer
No full textUrinary bladder cancer (BC) is the most common malignancy of the urinary tract, responsible for significant mortality and morbidity worldwide. In Europe, an estimate of 105.000 new cases of bladder cancer are diagnosed annually, while approximately 20.000 patients die from bladder cancer each year. Nearly all bladder cancers are carcinomas, arising from the urothelium. At presentation, 75-85% of tumours are restricted to the mucosa, or invade the lamina propria mucosae. The remainder present with invasion of the muscular layer of the bladder wall or extend to perivesical tissue, adjacent organs and even the pelvic wall. The behaviour of urothelial cell carcinoma (UCC) is highly diverse and defined by two separate, but related processes: tumour recurrence and progression. In specific, more than 60% of the superficial tumours will recur at least once and progress to less differentiated or invasive neoplasms in a significant percentage of patients. In current clinical practice the most useful prognostic parameters are tumour grade, stage, size, prior recurrence rate and the synchronous presence of carcinoma in situ (CIS). However, neither recurrence nor progression can be predicted accurately and a better understanding of the natural history of UCC is expected upon the elucidation its molecular mechanisms. Rho kinases comprise a major subgroup of the Ras superfamily that mediate housekeeping aspects of cell biology. In particular, Rho kinases play a fundamental role in the reorganization of actin and microtubule cytoskeleton. Therefore, Rho proteins govern a variety of cytoskeletondependent processes including alterations in cell shape, polarity, adhesion, cell motility, as well as cell to cell and cell to matrix interactions. Furthermore, Rho proteins appear to be involved in gene transcription by activating Serum Response Factor (SRF) and in the regulation of cell cycle progression, growth and apoptosis. Rho members affect the process of tumourigenesis either by over-expression of some members of the family with oncogenic activity or by down-modulation of other members with suggested tumour suppressor activity. Moreover, since Rho GTPases regulate actomyosin-based contractility and extracellular matrix remodelling, they may be involved in the transmigration of cells, thereby facilitating local invasiveness and metastasis. In the present study we investigated the expression of RhoA, RhoB, RhoC, Rac1 and Cdc42 kinases in UCC of the urinary bladder. Additionally, we determined the expression profile of 107 Rho-associated genes, including GTPases, GDIs, GAPs and GEFs. The expression was investigated using microarrays, qPCR and Western blotting in 77 UCC specimens compared to paired normal urothelium. Computational analysis was also performed on Gene Expression Omnibus datasets in order to further elucidate whether differences in the expression of Rho kinases occur among xviii superficial and invasive tumours. Further microarray analysis was carried out for the expression profiling of the Rho-associated genes. Finally, the correlation of the mRNA and protein levels of the genes of interest was also examined in UCC and normal tissue, using the Spearman rank correlation test According to our results RhoB RNA and protein levels were significantly lower in UCC, suggesting a tumour-suppressor role. On the contrary, RNA of RhoC and protein levels of RhoA, RhoC and Cdc42, respectively, were significantly higher in UCC compared to normal urothelium. High Cdc42 RNA levels correlated with worse overall survival, whereas high RhoB RNA levels correlated both with better overall and cancer-specific survival. Computational analysis verified the expression profile of Rho kinases among superficial UCC, muscle-invasive UCC and normal urothelium. Our findings in conjunction with the results of other investigators indicate that overexpression of either RhoA and/or RhoC is involved in tumourigenesis irrespective of its origin. In addition, over-expression of RhoA appears to be one of the events leading to more aggressive UCCs, characterized by higher histological grade and more extensive local invasion. RhoB in BC plays a tumour suppressor role, opposing the positive functions of RhoA and RhoC. Regarding its role in other malignancies a review of literature yielded contradictory results. These data suggest different Rho kinases play different roles in tumourigenesis and one Rho kinase may have different roles in different tumour development. Rac1 and Cdc42 seem to cross activate RhoA and/or RhoC in UCC and may contribute to tumour formation and progression. More importantly, preliminary evidence suggests that Cdc42 silencing leads to growth suppression in human bladder-cancer cells. In conclusion, the present study showed that the RNA and protein levels of RhoC and the protein levels of RhoA and Cdc42 were significantly higher in UCC compared to normal tissue. RhoB RNA and protein levels exhibited an inverse expression profile, supporting its role as a tumour suppressor gene. Elucidating the regulatory mechanisms that modulate Rho-mediated signaling pathways in tumours may therefore provide novel targets for small molecule therapeutic agents against cance
Mixed adenoneuroendocrine carcinoma of the urethra
No full textMixed adenoneuroendocrine carcinoma (MANEC) is a rare tumour found predominantly in the gastrointestinal tract. Comprising adenocarcinomatous and neuroendocrine components, MANEC have been reported in the bladder. We report the first case to our knowledge of a MANEC arising in the urethra. A 62-year-old woman presented with a suburethral mass. Initial excision of the mass revealed it to be a MANEC. Immunohistochemistry staining was positive for CK20 and synaptophysin associated with neuroendocrine tumours. Cross-sectional imaging ruled out metastases and the patient underwent radical urethrectomy, vaginal reconstruction and Mitrofanoff urinary diversion. The patient declined adjuvant chemotherapy and remained under regular surveillance. MANECs are uncommon tumours and treatment was therefore guided by expert opinion. A multidisciplinary approach is essential with the early involvement of surgeons, oncologists, histopathologist, radiologist and neuroendocrine specialists.</p
Spotlight on Differentially Expressed Genes in Urinary Bladder Cancer
No full textIntroduction: We previously identified common differentially expressed (DE) genes in bladder cancer (BC). In the present study we analyzed in depth, the expression of several groups of these DE genes. Materials and Methods: Samples from 30 human BCs and their adjacent normal tissues were analyzed by whole genome cDNA microarrays, qRT-PCR and Western blotting. Our attention was focused on cell-cycle control and DNA damage repair genes, genes related to apoptosis, signal transduction, angiogenesis, as well as cellular proliferation, invasion and metastasis. Four publicly available GEO Datasets were further analyzed, and the expression data of the genes of interest (GOIs) were compared to those of the present study. The relationship among the GOI was also investigated. GO and KEGG molecular pathway analysis was performed to identify possible enrichment of genes with specific biological themes. Results: Unsupervised cluster analysis of DNA microarray data revealed a clear distinction in BC vs. control samples and low vs. high grade tumors. Genes with at least 2-fold differential expression in BC vs. controls, as well as in non-muscle invasive vs. muscle invasive tumors and in low vs. high grade tumors, were identified and ranked. Specific attention was paid to the changes in osteopontin (OPN, SPP1) expression, due to its multiple biological functions. Similarly, genes exhibiting equal or low expression in BC vs. the controls were scored. Significant pair-wise correlations in gene expression were scored. GO analysis revealed the multi-facet character of the GOIs, since they participate in a variety of mechanisms, including cell proliferation, cell death, metabolism, cell shape, and cytoskeletal re-organization. KEGG analysis revealed that the most significant pathway was that of Bladder Cancer (p = 1.5x10(-31)). Conclusions: The present work adds to the current knowledge on molecular signature identification of BC. Such works should progress in order to gain more insight into disease molecular mechanisms
Incidence of needle‐tract seeding following prostate biopsy for suspected cancer: a review of the literature
No full textWorkplace personal exposure to respirable PM fraction: A study in sixteen indoor environments
No full textThe present paper focuses on respirable particulate matter (RPM) measurements conducted at the breathing zone of adult volunteers in sixteen different working environments: two offices, a house, a chemical laboratory, a non- smoking shop, a pharmacy store, a car garage, a hairdresser's store, a photocopy store, a taxi, a gym, a mall, a restaurant, a bar, a kiosk and a school. The sixteen different cases were categorized according to the location, the type of the activities taking place indoors, the number of occupants, the proximity to heavy traffic roads, the ventilation pattern etc. According to the results, the maximum particle concentration (in average 285 μg m-3) was recorded at the hairdresser store while the minimum concentration was measured in the cases of the housewife and the employee in the non-smoking shop (in average 30 μg m-3). The results indicated smoking as a factor which strongly influences the exposure levels of both smokers and passive smokers. Furthermore, it was found that the building ventilation pattern comprises an important factor influencing the exposure levels especially in cases of buildings with great number of visitors (resuspension) and smoking. © Author(s) 2014
Linear correlations in chromosomal-based gene expression in urinary bladder cancer
No full text<p>Introduction & Objectives: Gene expression is a very tidy and well coordinated procedure. Consecutive genes are often similarly expressed. We hypothesized that correlations might exist between genes of the same chromosome, yet belonging to different urinary bladder cancer (BC) samples, in order to indicate a common regulation for genes following this pattern.</p>
<p>Materials & Methods: We analyzed BC gene expression profiles, with emphasis in linear correlations of gene expression based on their chromosomal locations. Samples from 10 human BCs and 5 normal tissues were analyzed by whole genome microarrays, along with a computational approach, for their expression profiles. After raw data normalization and classification, differentially expressed genes (DE) were sorted according to their chromosome distributions and were further investigated for linear correlations among them. Chromosomal activity in terms of gene expression was measured by calculating the average expression of all DE genes for each chromosome, both for tumour and control samples.</p>
<p>Results: Chromosome-based expression analysis predicted that among the most active chromosomes were chromosomes 9 and X. Similarly, control samples also manifested high expression activity on the X chromosome. The genes that exhibited significant linear correlations (p<0.05) among tumor samples on chromosomes 4, 8, 13, 21 and 22, were as follows: TACR3, RNF150, ANXA10, CENTD1, EXOC1, GRSF1 for chromosome 4; ANXA13, DENND3, FGF20, EFHA2, DNAJC5B, MRPS28, FABP5 for chromosome 8; ITGBL1, RXFP2, KL, MYCBP2, FARP1 for chromosome 13; KRTAP19-1, IFNAR1, SON for chromosome 21; MORC2, PLA2G6, ACO2, ARHGAP8 for chromosome 22; SERPINA7, TMEM164, ARHGAP6, APLN, FHL1, PNMA6A, UBL4A, PRDX4, POLA1, MXRA5 for chromosome X.</p>
<p>Conclusions: Despite the fact that linear correlations occurred among distinct patients, the expression of the genes appeared to be correlated among them, in a similar manner. We have previously reported that there are hints of common mechanisms between BCs of different stage/grade, employing microarray analysis. Chromosomal correlation analysis comes to support our previous findings, since it revealed genes bearing common regulation among samples of different histology. Gene expression correlations can further assist us to understand more in-depth the mechanisms underlying tumour progression and biology.</p
Cross-platform comparisons of microarray data. Elucidation of common differentially expressed genes in bladder cancer.
No full text<p>INTRODUCTION: Parallel gene-expression monitoring is a powerful tool for analyzing relationships among tumors, discovering new tumor subgroups, assigning tumors to pre-defined classes, identifying co-regulated or tumor stage-specific genes and predicting disease outcome. Previous gene expression studies have focused on identifying differences between tumor samples of the same type.</p>
<p>AIM OF STUDY: Using a reverse engineering approach, we searched for common expression profiles among tumor samples. We analyzed the gene expression profile of bladder cancer (BC) and determined the differentially expressed (DE) genes between cancer and healthy tissue, using cross-platform comparisons.</p>
<p>MATERIALS AND METHODS: We performed cDNA microarray analysis, comprising both in-house experimental and publicly available GEO microarray data. In total, our pooled microarray analysis was composed of 17 control samples (n=5, for the CodeLink platform; and n=12, for the remaining microarray platforms) and 129 BC samples (n=10, for the CodeLink platform; and n=119, for the remaining microarray platforms). Tumor samples were separated into the following groups: Ta/T1 without CIS; Ta/T1 with CIS; Ta-grade 1; Ta-grade 3; T1-grade2; T1-grade 3; T2-grade 2-4. Each group was compared against all control samples and the DE genes were identified. Data were clustered with different algorithms.</p>
<p>RESULTS:</p>
<p>A two-sample T-test analysis for all tumor samples vs. all normal samples, revealed 434 DE genes between the two tissue groups. Hierarchical clustering (HCL) showed a clear distinction among tumor samples. In total, 17 genes appeared to be commonly expressed among all BC samples: BMP4, CRYGD, DBH, GJB1, KRT83, MPZ, NHLH1, TACR3, ACTC1, MFAP4, SPARCL1, TAGLN, TPM2, CDC20, LHCGR, TM9SF1 and HCCS. Three groups of genes were down-regulated in all samples: BMP4, CRYGD, DBH, GJB1, KRT83, MPZ, NHLH1, TACR3 in cluster 79; ACTC1, MFAP4, SPARCL1, TAGLN in cluster 81; and TPM2 in cluster 82. CDC20, TM9SF1 and HCCS appeared to be simultaneously over-expressed in all tumor groups. LHCGR was differentially expressed in 108/129 (83.7%) of the BC samples.</p>
<p>DISCUSSION: Through this investigation we were able to identify several important factors that warrant further investigation both as prognostic markers and as therapeutic targets. Such approaches may provide a better insight into tumorigenesis and tumor progression.</p
Spotlight on differentially expressed genes in urinary bladder cancer.
No full textINTRODUCTION: We previously identified common differentially expressed (DE) genes in bladder cancer (BC). In the present study we analyzed in depth, the expression of several groups of these DE genes. MATERIALS AND METHODS: Samples from 30 human BCs and their adjacent normal tissues were analyzed by whole genome cDNA microarrays, qRT-PCR and Western blotting. Our attention was focused on cell-cycle control and DNA damage repair genes, genes related to apoptosis, signal transduction, angiogenesis, as well as cellular proliferation, invasion and metastasis. Four publicly available GEO Datasets were further analyzed, and the expression data of the genes of interest (GOIs) were compared to those of the present study. The relationship among the GOI was also investigated. GO and KEGG molecular pathway analysis was performed to identify possible enrichment of genes with specific biological themes. RESULTS: Unsupervised cluster analysis of DNA microarray data revealed a clear distinction in BC vs. control samples and low vs. high grade tumors. Genes with at least 2-fold differential expression in BC vs. controls, as well as in non-muscle invasive vs. muscle invasive tumors and in low vs. high grade tumors, were identified and ranked. Specific attention was paid to the changes in osteopontin (OPN, SPP1) expression, due to its multiple biological functions. Similarly, genes exhibiting equal or low expression in BC vs. the controls were scored. Significant pair-wise correlations in gene expression were scored. GO analysis revealed the multi-facet character of the GOIs, since they participate in a variety of mechanisms, including cell proliferation, cell death, metabolism, cell shape, and cytoskeletal re-organization. KEGG analysis revealed that the most significant pathway was that of Bladder Cancer (p = 1.5×10(-31)). CONCLUSIONS: The present work adds to the current knowledge on molecular signature identification of BC. Such works should progress in order to gain more insight into disease molecular mechanisms
The tumour suppressor RhoB is an independent prognostic factor for metastasis in urinary bladder cancer.
No full text<p>Introduction & Objectives: Rho members may affect the process of tumourigenesis either by over-expression of some members of the family with oncogenic activity or by down-modulation of other members with suggested tumour suppressor activity.</p>
<p>Materials & Methods: Rho gene mRNA expression was studied in 77 bladder cancer (BC) specimens. The Kaplan-Meier method was used to estimate survival as a function of time, and survival differences were assessed by the Log-rank test. Logistic regression (univariate and multivariate) analysis was performed to determine the potential predictors of survival, recurrence and metastasis.</p>
<p>Results: The cases whose tumours exhibited increased levels of RhoB mRNA expression exhibited worse overall and cancer-specific survival rates, than those expressing decreased RhoB mRNA levels. Moreover, those cases whose tumours exhibited high Cdc42 mRNA expression showed a worse overall survival rate than those expressing low Cdc42 levels. High RhoC levels tended to correlate with better survival. Although univariate analysis, using the Cox proportional hazards model, showed that RhoB has a tendency for being an independent prognostic factor for overall survival (p=0.086), both univariate and multivariate analysis for RhoA, RhoC, Rac1 and Cdc42 did not exhibit the same tendency. Moreover, no gene was identified as independent prognostic factor for tumour recurrence. Finally, both univariate and multivariate analysis identified RhoB independent prognostic factors for metastasis (p=0.012 and p=0.050, respectively).</p>
<p>Conclusions: Our results confirm a tumour suppressor role for RhoB in bladder cancer, opposing the positive functions of RhoA and RhoC. Moreover, our analysis identified RhoB as an independent prognostic factor for metastasis.</p
