Association of FGFR alterations with FGFR 1-4 gene expression in TUR biopsies and matched NMP22 urine levels in early bladder cancer of the prospective real world clinico-pathological register trial: BRIDGister.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e16533-e16533
Author(s):  
Ralph M. Wirtz ◽  
Frank Friedersdorff ◽  
Elke Veltrup ◽  
Ergin Kilic ◽  
Richard Watts ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bladder Cancer ◽  
Mrna Expression ◽  
Real World ◽  
Standardized Test ◽  
Urine Samples ◽  
Tissue Samples ◽  
Basal Marker ◽  
Commercial Kits ◽  
Urine Levels

e16533 Background: The objective of the present study was to identify molecular charactersitics associated with FGFR positive tumors in matched urine and TUR biopsy samples from patients being suspicious of bladder cancer and undergoing first TURB at the pilot center of the multicentric BRIDGister Real World Experience trial that are helpful for patient management and prognosis. Methods: For this pilot study paraffin fixed pretreatment tissue samples from the first TURB of 28 pts participating in the BRIDGister trial and matched urine samples were prospectively collected and analyzed. RNA from FFPE tissues were extracted by commercial kits and analyzed by Therascreen FGFR IVD kit (Qiagen GmbH, Hilden). Relative gene expression of subtyping markers (KRT5, KRT0) as well as FGFR1, FGFR2, FGFR3, FGFR4, ERBB2 was centrally tested by standardized test systems (STRATIFYER Molecular Pathology GmbH, Cologne). In addition urine samples were analyzed by commercially available urine tests (UBC Rapid, BTA stat, NMP22). Spearman correlation, Kruskal-Wallis, MannWhitney and Sensitivity/Specificity tests were done by JMP 9.0.0 (SAS software). Results: The pilot cohort of the BRIDGister trial consisted of 28 patients (median age: 73, male 71% vs female 29%) of diverse clinical stages (Benign lesions/no tumor 21%, pTa 32%, pT1 21%, pT2 21%) and WHO 1973 grade (G1 7%, G2 43%, G3 21%). Based on FFPE tissue testing using Therascreen FGFR IVD kit 9 out of 28 patients exhibited FGFR alterations (32%). FGFR positive tumors were associated with high expression of FGFR3 mRNA (r = 5.951, p = 0.0011) but low FGFR1 mRNA as well as FGFR4 mRNA (r = -0.3882, p = 0.0412 and r = -0.6305, p = 0.0004, respectively). Interestingly, FGFR alteration was positively associated with the basal marker KRT5 (r = 0.3929, p = 0.0386), but not with luminal KRT20 mRNA expression (r = -0.0208, p = 0.9179). Moreover FGFR3 altered bladder cancer was associated with elevated NMP22 levels in pretreatment urine (r = 0.3978, p = 0.0361). Conclusions: In early bladder cancer FGFR3 alterations are tightly associated with a characteristic FGFR mRNA signature. Mutation/Fusion of FGFR3 results in high FGFR3 but low FGFR1 and FGFR4 mRNA expression, which might be i.a. relevant for the response to FGFR inhibition and important to predict outcome of FGFR inhibitors. Morever FGFR alteration was associated with elevated NMP22 urine levels, which might be helpful for detecting and monitoring FGFR altered bladder cancer in a non invasive fashion. These results warrant further investigation and its impact on outcome prediction (BCG responsiveness, recurrence, proegression, etc) will be prospectively analyzed in the framework of the ongoing multicenter BRIDGister Real World Experience trial.

FGFR testing from matched tissue and urine samples within the prospective real world clinico-pathological register trial BRIDGister.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e16532-e16532
Author(s):  
Ralph M. Wirtz ◽  
Richard Watts ◽  
Ronny Kellner ◽  
Reinhard Ortmann ◽  
Torsten Horns ◽  
...  
Keyword(s):  
Real World ◽  
Benign Lesion ◽  
Digital Pcr ◽  
Urine Samples ◽  
Urine Test ◽  
Tissue Samples ◽  
Before And After ◽  
Commercial Kits ◽  
Urine Testing ◽  
Matched Tissue

e16532 Background: The objective of the present study was to assess FGFR mutattions and fusions from matched urine and tissue samples from patients suspicious of bladder cancer and undergoing first TURB at the pilot center of the multicentric BRIDGister RealWorld Experience trial Methods: For this pilot study paraffin fixed pretreatment tissue samples from the first TURB of 28 pts participating in the BRIDGister trial and matched urine samples were prospectively collected and analyzed. RNA from FFPE tissues were extracted by commercial kits and analyzed by Therascreen FGFR IVD kit (Qiagen GmbH, Hilden). In addition urine samples were shipped for central isolation of extracellular vesicles and extraction of RNA (exoRNA.Exosome Diagnostics GmbH, Martinsried) and subsequently centrally analyzed by QIAcuity digital PCR (Qiagen, Hilden). Additional urine testing was performed by further technologies including central cytology. Concordance, Kruskal-Wallis, MannWhitney and Sensitivity/Specificity tests were analyzed by JMP 9.0.0 (SAS software). Results: The pilot cohort of the BRIDGister trial consisted of 28 patients (median age: 73, male 71% vs female 29%) of diverse clinical stages (Benign lesions/no tumor 21%, pTa 32%, pT1 21%, pT2 21%) and WHO 1973 grade (G1 7%, G2 43%, G3 21%). Based on FFPE tissue testing using Therascreen FGFR IVD kit 9 out of 28 patients exhibited FGFR alterations (32%). Based on exosomal RNA (exoRNA) and subsequent dPCR testing 8 out of 21 matched urine sampels were FGFR positive (38%). Comparison with tissue testing as probable gold standard revealed 71% sensitivity, 78% specificity, 63% PPV and 85%NPV. There were 3 patients being FGFR positive for exoRNA from urine with no mutation found in the corresponding TUR biopsy. One of these mutations could be validated by independent urine test. Furthermore one tumor harbored two tissue mutations (R248C, Y373C) but three urine mutations (R248C, Y373C, G370C) indicating substantial tumor heterogeneity. One FGFR3-TACC3 fusion was detected from a benign lesion, which was not found by the exosomal urine test. Conclusions: Extraction of exosomal RNA from urine followed by highly sensitive dPCR mutation testing is feasible with good concordance to matched tissue testing. Urine testing bears the potential of detecting additional mutations in a real world setting and might evolve as alternative approach for FGFR3 screening in a non invasive fashion without the need of transurethral biopsy. Discordant cases are further followed up and might reveal validation of mutation status in upcoming recurrences.Further exploration is warranted and includes the potential of monitoring patients with FGFR before and after therapeutic intervention. By the time of the congress an update of the data with approximately 50 matched pairs will be presented.

scholarly journals Prognostic value of CLIC3 mRNA overexpression in bladder cancer

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8348
Author(s):  
Mei Chen ◽  
Shufang Zhang ◽  
Xiaohong Wen ◽  
Hui Cao ◽  
Yuanhui Gao
Keyword(s):  
Gene Expression ◽  
Bladder Cancer ◽  
Mrna Expression ◽  
Prognostic Value ◽  
Multivariate Analyses ◽  
Gene Set Enrichment Analysis ◽  
The Cancer Genome Atlas ◽  
Functional Enrichment ◽  
Receptor Interaction ◽  
Normal Tissues

Background Human intracellular chloride channel 3 (CLIC3) is involved in the development of various cancers, but the expression and prognostic value of CLIC3 mRNA in bladder cancer (BC) remain unclear. Methods The gene expression data and clinical information of CLIC3 were obtained from the Gene Expression Omnibus (GEO) database and verified in the Oncomine and The Cancer Genome Atlas (TCGA) database. The expression of CLIC3 mRNA in BC tissues and adjacent normal tissues was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The Kaplan-Meier method was used to analyze the relationship between the expression of CLIC3 mRNA and the prognosis of BC. Cox univariate and multivariate analyses were performed on the overall survival and tumor-specific survival of BC patients. The genes coexpressed with CLIC3 were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). CLIC3-related signal transduction pathways in BC were explored with gene set enrichment analysis (GSEA). Results The expression of CLIC3 mRNA in BC tissues was higher than that in normal tissues (P < 0.01). High CLIC3 mRNA expression was associated with age (P = 0.021) and grade (P = 0.045) in BC patients. High CLIC3 mRNA expression predicted a poor prognosis in BC patients (P < 0.05). Cox univariate and multivariate analyses showed that high CLIC3 mRNA expression was associated with tumor-specific survival in BC patients (P < 0.05). Functional enrichment analyses indicated that CLIC3 may be significantly associated with the cell cycle, focal adhesion, the extracellular matrix (ECM) receptor interaction and the P53 signaling pathway. Conclusions CLIC3 mRNA is highly expressed in BC, and its high expression is related to the adverse clinicopathological factors and prognosis of BC patients. CLIC3 can be used as a biomarker for the prognosis of BC patients.

Assessment of the diagnostic value of microsatellite instability in the urine of bladder cancer patients.

2019 ◽  
Vol 37 (7_suppl) ◽  
pp. 384-384
Author(s):  
Hussein Mustafa Khaled ◽  
Abdel-Rahman Zekri ◽  
Mai B Mohamed ◽  
Fatma M Diab ◽  
Mona Abdellateif ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Microsatellite Instability ◽  
Tumor Tissue ◽  
High Sensitivity ◽  
Diagnostic Value ◽  
Urine Samples ◽  
Tissue Samples ◽  
Predictive Values ◽  
Promising Tool ◽  
Sensitivity Specificity

384 Background: Microsatellite alterations in urine sediments have proved to be a promising tool for detection of bladder cancer (BC) due to its high sensitivity and specificity. Methods: We assessed the possible prognostic and predictive values of microsatellite alterations in tissue samples and urine sediments obtained from Egyptian patients with BC, and their utility as diagnostic, prognostic and predictive value. Microsatellite instability (MSI) and loss of heterozygosity (LOH) were assessed using 13 microsatellite markers in tumor tissue and urine sediments of 30 patients with BC. The concordance between MSI in tissue and urine samples was determined. Results: We found that MSI was more frequent than LOH (100% and 46.7%; respectively). D16S310, MBP and IFN-α showed the highest MSI frequency in urine samples (70%, 70% and 66.67%; respectively), while MBP, ACTBP2 and D9S171 (66.67%, 63.33%, and 60%; respectively) were the most frequently detected in tumor tissue. All markers correlated significantly with the pathological subtypes (more frequent in TCC) and hematuria. The concordance between tissue and urine was statistically significant for , D9S171, D16S476, FGA and ACTBP2 (P = 0.04, 0.015, 0.02 and 0.007; respectively). When we combined D16S476 and D9S171, the sensitivity, specificity, PPV and NPV reached (80.0%, 75.0%, 82.8% and 71.4%; respectively) for diagnosis of BC . Conclusions: Thus MSI in urine sediments could be a sensitive and reliable method for diagnosis of BC.

PSI-5 Changes in the expression of MyD88, TRAF6, NFKB1, and NFKB1a on digestive tracts of 0 to 42 days-old calves from the tropics of Mexico

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 283-283
Author(s):  
Mariana Álvarez Pérez ◽  
Carolina Robles-Rodriguez ◽  
Laura González Dávalos ◽  
Armando Shimada ◽  
Alfredo Varela ◽  
...  
Keyword(s):  
Gene Expression ◽  
Immune Response ◽  
Mrna Expression ◽  
Digestive Tract ◽  
Adaptive Immune Response ◽  
Innate Immune ◽  
Tissue Samples ◽  
Adaptive Immune ◽  
Enteric Diseases ◽  
The Tropics

Abstract During the first and second weeks of life, calves are extremely susceptible to enteric diseases, their digestive tract is developing and their thermoregulation is not adequate; these being some factors that can contribute to the lack of their weight gain (Hulbert & Moisá, 2016). In addition, they have a reduced capacity to generate an innate and adaptive immune response, in other words, they are still not immunocompetent (Costa et al., 2017; Hulbert & Moisá, 2016), since they are born with all the essential immune components but many of them are not functional until calves are at least 2 to 4 weeks old (Chase et al., 2008). With the aim to identify the development of the innate immune response through the expression of MyD88, TRAF6, NFKB1, and NFKB1a mRNA, in different ages and regions of the digestive tract, qPCR tests were held. Twelve Zebu crossbred calves of 0, 7, 28, and 42 days of age were sampled (three animals of each age). Tissue samples included: rumen, duodenum, and ileum. Gene expression was determined by quantitative PCR (qPCR). Changes in the expression of TRAF6, NFKB1 and NFKB1a mRNA were different in rumen and ileum (P &lt; 0.05) at day 28 of age. MYD88 mRNA expression was different only in ileum (P &lt; 0.05). MyD88, TRAF6, NFKB1, and NFKB1a mRNA expression of rumen tended to decrease and in the case of ileum to increase with age; however, this was until day 28 and not in the first week of life, as mentioned by some other authors.

Quantitative Analysis of Heparanase Gene Expression in Normal Cervical, Cervical Intraepithelial Neoplastic, and Cervical Carcinoma Tissues

2009 ◽  
Vol 19 (9) ◽  
pp. 1614-1619 ◽  
Author(s):  
Eugene Varchalama ◽  
Alexander Rodolakis ◽  
Areti Strati ◽  
Theocharis Papageorgiou ◽  
Christos Valavanis ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cervical Cancer ◽  
Heparan Sulfate ◽  
Mrna Expression ◽  
Tumor Size ◽  
Invasive Cervical Cancer ◽  
Cancer Tissue ◽  
Low Grade ◽  
Tissue Samples ◽  
Cervical Cancer Tissue

Heparanase is an endoglycosidase that specifically cleaves heparan sulfate side chains of heparan sulfate proteoglycans, the major proteoglycans in the extracellular matrix and cell surfaces. Traditionally, heparanase activity was implicated in cellular invasion associated with angiogenesis, inflammation, and cancer metastasis. More recently, heparanase up-regulation was documented in an increasing number of primary human tumors. Ιn this study, we sought to investigate the expression of heparanase messenger RNA (mRNA) in normal cervical tissue and intraepithelial cervical lesion and its clinicopathologic importance in invasive cervical cancer. Gene expression of heparanase was assessed by quantitative real-time reverse transcriptase polymerase chain reaction in 28 normal cervical, 26 intraepithelial neoplastic, and 48 cervical cancer tissue samples. Heparanase mRNA expression was different between the 3 groups and lower in normal cervical specimens in relationship with intraepithelial cervical lesions and invasive cervical cancer tissue samples (P = 0.048). Gradually increasing expression of heparanase was evident as the cells progressed from low-grade to high-grade squamous intraepithelial lesions (P = 0.002). In invasive cervical cancer cases, there was a direct correlation between heparanase expression and tumor size (P = 0.002). In cases treated with radical hysterectomy and pelvic lymphadenectomy, the heparanase mRNA expression was significantly higher in tumors exhibiting lymph vascular space invasion (P = 0.044) and in cases with big tumor size (P = 0.005). In our study, we did not find any significant correlation between disease-free and overall survival rates and expression of heparanase (P = 0.396 and P = 0.712, respectively). The results of this study suggest that the gene expression of heparanase in cervical cancer enhances growth, invasion, and angiogenesis of the tumor and may have therapeutic applications.

Association of KRT20 and KRT5 with response to neoadjuvant chemotherapy in patients diagnosed with muscle invasive bladder cancer.

2020 ◽  
Vol 38 (6_suppl) ◽  
pp. 562-562
Author(s):  
Florian Roghmann ◽  
Moritz Reike ◽  
Ralph Wirtz ◽  
Maximilian Kriegmair ◽  
Philipp Erben ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Neoadjuvant Chemotherapy ◽  
Mrna Expression ◽  
Expression Patterns ◽  
High Risk Group ◽  
Invasive Bladder Cancer ◽  
Study Cohort ◽  
Muscle Invasive Bladder Cancer ◽  
Tissue Samples ◽  
Muscle Invasive

562 Background: Patients with muscle-invasive bladder cancer (MIBC) that underwent neoadjuvant chemotherapy (NAC) prior to radical cystectomy (RC) show improved overall survival. Those with a pathological complete response (pCR) usually have the best prognosis. In the literature, improved response to NAC has been associated with basal tumor characteristics in MIBC so far. The aim of the present study was to examine the association of luminal (KRT20) and basal (KRT5) mRNA expression patterns at transurethral resection (TUR) with pCR at RC after NAC in a contemporary cohort of consecutive MIBC patients. Methods: Clinical Data and formalin fixed paraffin embedded tumor tissue samples from TUR and RC of 49 patients with MIBC were retrospectively analyzed. Using RT-PCR KRT20 and KRT5 mRNA expression were measured in 40-∆Ct values and normalized against the control gene CALM2. Statistical analyses comprised nonparametric and chi2 testing, partition models and spearman correlation analyses. Results: The study cohort had a median age of 63 years and consisted of 38/49 (78%) males. After NAC, 17/49 (35%) patients had cPR. Using partition models, we found that patients with high-KRT20 (≥39.5 ∆Ct) had a higher chance of pCR (57% vs. 26%, p=0.04). Using a cutoff for KRT5 at <38.1 ∆Ct within the subgroup of patients with low-KRT20 (<39.5 ∆Ct, n=35), we found poorest response among low-KRT20/low-KRT5 compared to low-KRT20/high-KRT5 and high-KRT20 (13% vs. 37% vs. 57%, p=0.29), respectively. For low-KRT20/low-KRT5, low-KRT20/high-KRT5 and high-KRT20 median KRT5 was 34.8 vs. 39.5 vs. 34.1 ∆Ct ( p=0.001) and median KRT20 was 37.9 vs. 32.9 vs. 40.1 ∆Ct,( p=0.001), respectively. Conclusions: Patients with MIBC showing high expression of KRT20 were more likely to show pCR at RC after NAC. Moreover, we were able to identify a high risk group of patients with lowKRT20/lowKRT5 that was less likely to achieve pCR at RC after NAC. Our findings are contradicting previous studies and need further verification in larger cohorts. However, our results might be useful for treatment stratification in MIBC patients.

scholarly journals A pilot study of urinary microRNA as a biomarker for urothelial cancer

2013 ◽  
Vol 7 (1-2) ◽  
pp. 28 ◽  
Author(s):  
Jaime Snowdon ◽  
Sandy Boag ◽  
Harriet Feilotter ◽  
Jason Izard ◽  
D. Robert Siemens
Keyword(s):  
Bladder Cancer ◽  
Pilot Study ◽  
Urothelial Cancer ◽  
Bladder Tumour ◽  
Fold Increase ◽  
Urine Samples ◽  
Tumour Resection ◽  
Cancer Tissue ◽  
High Grade ◽  
Tissue Samples

Objective: MicroRNAs (miRNAs) are part of a class of small ribonucleic acid (RNAs). They are important regulatory molecules, involved in several cell processes, such as developmental timing, stem cell division and apoptosis. Dysregulated miRNAs have been identified in several human malignancies, including bladder cancer tissue samples, and may confer a “tumour signature” that can be exploited for diagnostic purposes. We report on a prospective pilot study investigating the diagnostic capability of miRNAs in the urine of patients with urothelial cancer.Methods: Voided urine samples were collected from patients with urothelial carcinoma just prior to bladder tumour resection, as well as age-matched healthy control patients. Pathology demonstrated both low- and high-grade cancer. Total RNA was isolated and quantitative reverse transcriptase-polymerase chain reaction was performed on the RNA extracts using primers for 4 miRNAs shown previously to be dysregulated in solid urothelial carcinomas with RNU6B as the endogenous control. Standard urine cytology was performed on all samples in a blinded fashion.Results: Two miRNAs of interest were dysregulated in the urine from cancer patients with miR-125b showing an average 10.42-fold decrease (p < 0.01) and miR-126 showing an average 2.70-fold increase (p = 0.30) in the cancer samples compared to the normal controls. The sensitivity and specificity of the cytology on the same urine samples were 50% and 80%, respectively. Using these 2miRNAs only, a decision-tree prediction model was generated for a validation cohort of patients yielding a specificity of 100% and a sensitivity of 80%.Discussion: This preliminary study of candidate urinary miRNA inpatients with low- and high-grade urothelial cancer demonstrated a significantly improved diagnostic accuracy over cytology. These results provide rationale for further studies on discovery and validation of candidate miRNAs in voided urine and may potentially lead to the development of a non-invasive and sensitive test for bladder cancer diagnosis and prognosis.

Identification of Suitable Internal Control Genes for Quantitative Real-Time PCR Expression Analyses in Peanut (Arachis hypogaea)

2010 ◽  
Vol 37 (1) ◽  
pp. 12-19 ◽  
Author(s):  
Yael Brand ◽  
Ran Hovav
Keyword(s):  
Gene Expression ◽  
Mrna Expression ◽  
Real Time ◽  
Internal Control ◽  
Real Time Pcr ◽  
Reference Genes ◽  
Developmental Stages ◽  
Peanut Kernel ◽  
Tissue Samples ◽  
The Stability

Abstract Real-time qPCR is currently the most sensitive technique available for the detection of low-level mRNA expression. For more reliable and precise gene expression analyses, real-time PCR data for a sequence of interest must be normalized against that of a control gene, which is uniformly expressed in various tissues and during different phases of development. So far, suitable internal controls for gene expression studies in peanut have not been identified. We assessed the expression of 10 frequently used housekeeping genes, specifically ubq10, gapdh, hel1, yls8, 14-3-3, 60s, ubc, ef-1α, act7, and adh3. Using the algorithms available through the GeNorm and NormFinder programs, the stability of their expression was estimated in a set of five diverse peanut tissue samples derived from a Virginia-type peanut cultivar (Shulamit). Collectively, the gene with the most stable expression across all of the examined tissues and both programs was adh3, followed by 60s and yls8, which had minimal estimated intra- and inter-tissue variation. The stability of two stable reference genes (adh3 and yls8) compared with two less stable (14-3-3 and ubq10) reference genes was validated in unpooled tissue samples from five peanut kernel developmental stages. Finally, the effect of the use of one or more reference genes on the observed relative expression levels of an important seed oil metabolism gene, diacylglycerol acyltransferase 1 (Dgat1), during kernel development was demonstrated. Based on findings, the suggestion is that adh3, or a combination of this gene with 60s and yls8 should be considered for use in quantitative mRNA expression analyses in Arachis, particularly in studies involving seed development; whereas ubq10 and gapdh should be avoided.

scholarly journals Gene Expression, DNA Methylation and Prognostic Significance of DNA Repair Genes in Human Bladder Cancer

2017 ◽  
Vol 42 (6) ◽  
pp. 2404-2417 ◽  
Author(s):  
Anita Wojtczyk-Miaskowska ◽  
Malgorzata Presler ◽  
Jerzy Michajlowski ◽  
Marcin Matuszewski ◽  
Beata Schlichtholz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
Bladder Cancer ◽  
Dna Repair ◽  
Mrna Expression ◽  
Promoter Methylation ◽  
Methylation Status ◽  
Prognostic Significance ◽  
Muscle Invasive ◽  
Repair Genes

Background/Aims: This study investigated the gene expression and DNA methylation of selected DNA repair genes (MBD4, TDG, MLH1, MLH3) and DNMT1 in human bladder cancer in the context of pathophysiological and prognostic significance. Methods: To determine the relationship between the gene expression pattern, global methylation and promoter methylation status, we performed real-time PCR to quantify the mRNA of selected genes in 50 samples of bladder cancer and adjacent non-cancerous tissue. The methylation status was analyzed by methylation-specific polymerase chain reaction (MSP) or digestion of genomic DNA with a methylation-sensitive restriction enzyme and PCR with gene-specific primers (MSRE-PCR). The global DNA methylation level was measured using the antibody-based 5-mC detection method. Results: The relative levels of mRNA for MBD4, MLH3, and MLH1 were decreased in 28% (14/50), 34% (17/50) and 36% (18/50) of tumor samples, respectively. The MBD4 mRNA expression was decreased in 46% of non-muscle invasive tumors (Ta/T1) compared with 11% found in muscle invasive tumors (T2-T4) (P<0.003). Analysis of mRNA expression for TDG did not show any significant differences between Ta/T1 and T2-T4 tumors. The frequency of increased DNMT1 mRNA expression was higher in T2-T4 (52%) comparing to Ta/T1 (16%). The overall methylation rates in tumor tissue were 18% for MBD4, 25% for MLH1 and there was no evidence of MLH3 promoter methylation. High grade tumors had significantly lower levels of global DNA methylation (P=0.04). There was a significant association between shorter survival and increased expression of DNMT1 mRNA (P=0.002), decreased expression of MLH1 mRNA (P=0.032) and the presence of MLH1 promoter methylation (P=0.006). Conclusion: This study highlights the importance of DNA repair pathways and provides the first evidence of the role of MBD4 and MLH3 in bladder cancer. In addition, our findings suggest that DNMT1 mRNA and MLH1 mRNA expression, as well as the status of MLH1 promoter methylation, are attractive prognostic markers in this pathology.

scholarly journals Identification of cell division cycle 20 as a candidate biomarker and potential therapeutic target in bladder cancer using bioinformatics analysis

2020 ◽  
Vol 40 (7) ◽  
Author(s):  
Peilin Shen ◽  
Xuejun He ◽  
Lin Lan ◽  
Yingkai Hong ◽  
Mingen Lin
Keyword(s):  
Gene Expression ◽  
Bladder Cancer ◽  
Cell Division ◽  
Mrna Expression ◽  
Therapeutic Target ◽  
Gene Expression Omnibus ◽  
Cell Division Cycle ◽  
Hub Genes ◽  
Potential Therapeutic Target ◽  
Protein Protein Interaction

Abstract Purpose: As bladder cancer (BC) is very heterogeneous and complicated in the genetic level, exploring genes to serve as biomarkers and therapeutic targets is practical. Materials and methods: We searched Gene Expression Omnibus (GEO) and downloaded the eligible microarray datasets. After intersection analysis for identified differentially expressed genes (DEGs) of included datasets, overlapped DEGs were identified and subsequently analyzed with Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Protein–Protein Interaction (PPI) and hub genes identification. Hub genes were further analyzed with mRNA expression comparation in Oncomine and Gene Expression Profiling Interactive Analysis (GEPIA) database, proteomics-based validation in The Human Protein Atlas (THPA) and survival analysis in GEO and Oncolnc database. Results: We analyzed five eligible GEO datasets and identified 76 overlapped DEGs mapped into PPI network with 459 edges which were mainly enriched in cell cycle pathway and related terms in GO and KEGG analysis. Among five identified hub genes, which are Cyclin-Dependent Kinase 1 (CDK1), Ubiquitin-Conjugating Enzyme E2 C (UBE2C), Cell Division Cycle 20 (CDC20), Microtubule Nucleation Factor (TPX2) and Cell Division Cycle Associated 8 (CDCA8); CDC20 and CDCA8 were confirmed as significant in mRNA expression comparation and proteomics-based validation. However, only CDC20 was considered prognostically significant in both GEO and Oncolnc database. Conclusions: CDC20 and CDCA8 were identified as candidate diagnostic biomarkers for BC in the present study; however, only CDC20 was validated as prognostically valuable and may possibly serve as a candidate prognostic biomarker and potential therapeutic target. Still, further validation studies are essential and indispensable.

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