scholarly journals Application of SPRi Biosensors for Determination of 20S Proteasome and UCH-L1 Levels in the Serum and Urine of Transitional Bladder Cancer Patients

2021 ◽  
Vol 11 (17) ◽  
pp. 7835
Author(s):  
Anna Sankiewicz ◽  
Tomasz Guszcz ◽  
Ewa Gorodkiewicz
Keyword(s):  
Bladder Cancer ◽  
Cancer Progression ◽  
Transitional Cell ◽  
Ubiquitin Proteasome System ◽  
Clinicopathological Parameters ◽  
Control Group ◽  
20S Proteasome ◽  
Ubiquitin Proteasome ◽  
Serum And Urine

The ubiquitin–proteasome system (UPS) participates in the degradation of proteins which play an important role in regulating the cell cycle, apoptosis, and angiogenesis, as well as in the immune system. These processes are important in carcinogenesis. Transitional cell carcinoma (TCC) is one of the predominant types of bladder cancer. The relationship between the ubiquitin–proteasome system and cancer progression has become a topic of increasing interest among researchers. In this work, we propose an application of surface plasmon resonance imaging (SPRi)-based biosensors for the detection of 20S proteasome and ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) in the blood serum and urine of patients with TCC. The aim of the study was to determine 20S proteasome and UCH-L1 concentrations and to correlate the results with clinicopathological parameters. The group of subjects consisted of 82 patients with confirmed TCC, in addition to a control group of 27 healthy volunteers. It was found that 20S proteasome and UCH-L1 concentrations were significantly elevated in both the serum and urine of TCC patients, compared with the healthy subjects. There was a correlation between 20S proteasome concentrations in serum and urine, as well as between serum proteasome and UCH-L1 concentration. The SPRi biosensor sensitive to 20S proteasome using PSI inhibitor as the receptor, and the SPRi biosensor sensitive to the UCH-L1 protein using the protein-specific antibody as the receptor is suitable for the determination of 20S proteasome and UCH-L1 in body fluids and can serve as useful tools in the investigation of cancer biomarkers.

Theoretical Studies of the Acid-Base Equilibria in a Model Active Site of the Human 20S Proteasome

2020 ◽  
Author(s):  
Jon Uranga ◽  
Lukas Hasecke ◽  
Jonny Proppe ◽  
Jan Fingerhut ◽  
Ricardo A. Mata
Keyword(s):  
Active Site ◽  
Boronic Acid ◽  
Computational Study ◽  
Ubiquitin Proteasome System ◽  
Bayesian Optimization ◽  
Inhibition Mechanism ◽  
20S Proteasome ◽  
Acid Base ◽  
Ubiquitin Proteasome ◽  
Infection Cycles

The 20S Proteasome is a macromolecule responsible for the chemical step in the ubiquitin-proteasome system of degrading unnecessary and unused proteins of the cell. It plays a central role both in the rapid growth of cancer cells as well as in viral infection cycles. Herein, we present a computational study of the acid-base equilibria in an active site of the human proteasome, an aspect which is often neglected despite the crucial role protons play in the catalysis. As example substrates, we take the inhibition by epoxy and boronic acid containing warheads. We have combined cluster quantum mechanical calculations, replica exchange molecular dynamics and Bayesian optimization of non-bonded potential terms in the inhibitors. In relation to the latter, we propose an easily scalable approach to the reevaluation of non-bonded potentials making use of QM/MM dynamics information. Our results show that coupled acid-base equilibria need to be considered when modeling the inhibition mechanism. The coupling between a neighboring lysine and the reacting threonine is not affected by the presence of the inhibitor.

Ubiquitin-proteasome System Is a Promising Target for Killing Cisplatin-resistant Bladder Cancer Cells

2021 ◽  
Vol 41 (6) ◽  
pp. 2901-2912
Author(s):  
KAZUKI OKUBO ◽  
MAKOTO ISONO ◽  
TAKAKO ASANO ◽  
NINA REßING ◽  
WOLFGANG A. SCHULZ ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Cancer Cells ◽  
Ubiquitin Proteasome System ◽  
Promising Target ◽  
Bladder Cancer Cells ◽  
Ubiquitin Proteasome

An importance of matrix-metalloprotease determination in urine of patients with bladder cancer

2021 ◽  
Vol 1 (1) ◽  
pp. 30-32
Author(s):  
Vladimir N. Tkachuk ◽  
Adel S. Al-Shukri ◽  
Dmitriy I. Danilchenko
Keyword(s):  
Bladder Cancer ◽  
Control Group ◽  
Depth Of Invasion ◽  
Matrix Metalloprotease ◽  
Cancer Occurrence ◽  
Bladder Diseases ◽  
Neoplastic Process ◽  
Group A ◽  
Experimental Group

The aimof this work is to estimate an influence of a depth and an extent of differentiation of neoplastic process on 2 and 9 matrix-metalloprotease levels in urine of patients with bladder cancer. Patients and methods.71 patients with bladder cancer have been investigated. 14 patients with chronic cystitis were included in an experimental group and 45 patients without bladder diseases were included in a control group, patients without urinary tract obstructions and infections were included in a control group. A level of metalloprotease was identified by a cimographic method. Results.It has been proved that enzymes MMP-2 and MMP-9 of urine proteolysis are high sensitive markers of bladder cancer occurrence and their concentration depends on a depth of invasion and an extent of tumor differentiation, and shows a disease course prognosis. Conclusion.It is reasonably to identify MMP-2 and MMP-9 of urine by the cimographic method for diagnostics and noninvasive determination of a depth of invasion and an extent of bladder cancer differentiation.

scholarly journals Involvement of the Ubiquitin-Proteasome System in the Formation of Experimental Postsurgical Peritoneal Adhesions

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Clara Di Filippo ◽  
Pasquale Petronella ◽  
Fulvio Freda ◽  
Marco Scorzelli ◽  
Marco Ferretti ◽  
...  
Keyword(s):  
Protein Degradation ◽  
Ubiquitin Proteasome System ◽  
20S Proteasome ◽  
Intracellular Protein ◽  
Peritoneal Adhesions ◽  
Proteasome Subunit ◽  
Peritoneal Tissue ◽  
Intracellular Protein Degradation ◽  
Ubiquitin Proteasome

We investigated the Ubiquitin-Proteasome System (UPS), major nonlysosomal intracellular protein degradation system, in the genesis of experimental postsurgical peritoneal adhesions. We assayed the levels of UPS within the adhered tissue along with the development of peritoneal adhesions and used the specific UPS inhibitor bortezomib in order to assess the effect of the UPS blockade on the peritoneal adhesions. We found a number of severe postsurgical peritoneal adhesions at day 5 after surgery increasing until day 10. In the adhered tissue an increased values of ubiquitin and the 20S proteasome subunit, NFkB, IL-6, TNF-αand decreased values of IkB-beta were found. In contrast, bortezomib-treated rats showed a decreased number of peritoneal adhesions, decreased values of ubiquitin and the 20S proteasome, NFkB, IL-6, TNF-α, and increased levels of IkB-beta in the adhered peritoneal tissue. The UPS system, therefore, is primarily involved in the formation of post-surgical peritoneal adhesions in rats.

scholarly journals Determination of ubiquitin fitness landscapes under different chemical stresses in a classroom setting

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
David Mavor ◽  
Kyle Barlow ◽  
Samuel Thompson ◽  
Benjamin A Barad ◽  
Alain R Bonny ◽  
...  
Keyword(s):  
Fitness Landscape ◽  
Ubiquitin Proteasome System ◽  
First Year ◽  
Graduate Curriculum ◽  
Specific Effects ◽  
C Terminus ◽  
Chemically Induced ◽  
Sequencing Studies ◽  
Ubiquitin Proteasome

Ubiquitin is essential for eukaryotic life and varies in only 3 amino acid positions between yeast and humans. However, recent deep sequencing studies indicate that ubiquitin is highly tolerant to single mutations. We hypothesized that this tolerance would be reduced by chemically induced physiologic perturbations. To test this hypothesis, a class of first year UCSF graduate students employed deep mutational scanning to determine the fitness landscape of all possible single residue mutations in the presence of five different small molecule perturbations. These perturbations uncover 'shared sensitized positions' localized to areas around the hydrophobic patch and the C-terminus. In addition, we identified perturbation specific effects such as a sensitization of His68 in HU and a tolerance to mutation at Lys63 in DTT. Our data show how chemical stresses can reduce buffering effects in the ubiquitin proteasome system. Finally, this study demonstrates the potential of lab-based interdisciplinary graduate curriculum.

scholarly journals New Electrochemiluminescent Immunoassay for the Determination of CYFRA 21-1: Analytical Evaluation and Clinical Diagnostic Performance in Urine Samples of Patients with Bladder Cancer

1999 ◽  
Vol 45 (11) ◽  
pp. 1944-1953 ◽  
Author(s):  
Marta Sánchez-Carbayo ◽  
Antonia Espasa ◽  
Virtudes Chinchilla ◽  
Enrique Herrero ◽  
Julián Megías ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Roc Analysis ◽  
Threshold Value ◽  
Urine Samples ◽  
Cytokeratin 19 ◽  
Analytical Evaluation ◽  
Clinical Value ◽  
Sensitivity Measurement ◽  
Serum And Urine

Abstract Background: A new electrochemiluminescent immunoassay (ECLIA) has been developed for the determination of cytokeratin 19 (CYFRA 21-1) in the Elecsys 2010 immunoassay system. Urinary CYFRA 21-1 might have a role in the diagnosis of bladder cancer. Methods: We performed an analytical evaluation of the CYFRA 21-1 ECLIA for serum and urine samples. The clinical value of urinary CYFRA 21-1 for the detection of bladder cancer was evaluated through its measurement in 226 urine samples from symptomatic and asymptomatic controls. Results: At concentrations of 2–30 μg/L, within-assay imprecision (CV) was below 2.1% for sera and 3.3% for urines, with interassay CVs below 3.3% for sera and 4.9% for urines. The day-to-day CV was <20% at concentrations >0.2 μg/L (functional sensitivity). Measurement of diluted samples showed that the assay estimated CYFRA 21-1 between 98% and 103% for sera and 98% and 105% for urines. Recovery of added CYFRA 21-1 was 99–105% for sera and 96–115% for urines. We separately compared serum and urine CYFRA 21-1 ECLIA results with those obtained with an IRMA (CIS bio international). Regression analysis for sera was: CYFRA 21-1 (ECLIA) = 0.520 + 1.018 CYFRA 21-1 (IRMA); [95% confidence interval (CI) (y-intercept), −0.260 to 1.309]; 95% CI (slope), 0.978–1.060; n = 100; Sy|x = 3.242; r2 = 0.987. For urine samples it was: CYFRA 21-1 (ECLIA) = 0.716 + 0.966 CYFRA 21-1 (IRMA); 95% CI (y-intercept), 0.009–1.422; 95% CI (slope), 0.956–0.976; n = 100; Sy|x = 4.136; r2 = 0.986. In urine samples voided by patients with and without bladder cancer, the best ROC analysis discrimination provided 81.0% (95% CI, 72.7–87.7%) sensitivity and 97.2% (95% CI, 90.2–99.6%) specificity at a threshold value of 5.7 μg/L. Conclusions: Our initial evaluation showed reliable analytical performance for urinary CYFRA 21-1, which might assist urologists in the detection of bladder cancer as a noninvasive adjunct to cystoscopy.

scholarly journals Biomarkers in Bladder Cancer: Present Status and Perspectives

2007 ◽  
Vol 2 ◽  
pp. 117727190700200 ◽  
Author(s):  
Wun-Jae Kim ◽  
Soongang Park ◽  
Yong-June Kim
Keyword(s):  
Gene Expression ◽  
Cancer Research ◽  
Bladder Cancer ◽  
Cancer Progression ◽  
Transitional Cell ◽  
Cancer Recurrence ◽  
Global Gene Expression ◽  
Cancer Therapies ◽  
Predictive Capacity ◽  
Specificity And Sensitivity

Bladder cancers are a mixture of heterogeneous cell populations, and numerous factors are likely to be involved in dictating their recurrence, progression and the patient's survival. For any candidate prognostic marker to have considerable clinical relevance, it must add some predictive capacity beyond that offered by conventional clinical and pathologic parameters. Here, the current situation in bladder cancer research with respect to identification of suitable prognostic markers is reviewed. A number of individual molecular markers that might predict bladder cancer recurrence and progression have been identified but many are not sufficiently sensitive or specific for the whole spectrum of bladder cancer diseases seen in routine clinical practice. These limitations have led to interest in other molecular parameters that could enable more accurate prognosis for bladder cancer patients. Of particular interest is the epigenetic silencing of tumor suppressor genes. Since the methylation of these genes can correlate with a poor prognosis, the methylation profile may represent a new biomarker that indicates the risk of transitional cell carcinoma development. In addition, bladder cancer research is likely to be revolutionized by high-throughput molecular technologies, which allow rapid and global gene expression analysis of thousands of tumor samples. Initial studies employing these technologies have considerably expanded our ability to classify bladder cancers with respect to their survivability. Future microarray analyses are likely to reveal particular gene expression signatures that predict the likelihood of bladder cancer progression and recurrence, as well as patient's survival and responsiveness to different anti-cancer therapies, with great specificity and sensitivity.

The ubiquitin–proteasome system and cancer

2005 ◽  
Vol 41 ◽  
pp. 187-203 ◽  
Author(s):  
Anny Devoy ◽  
Tim Soane ◽  
Rebecca Welchman ◽  
R. John Mayer
Keyword(s):  
Cell Cycle ◽  
Protein Phosphorylation ◽  
Cancer Progression ◽  
Kinase Inhibitors ◽  
Cell Transformation ◽  
Ubiquitin Proteasome System ◽  
Regulatory Pathways ◽  
Regulatory Processes ◽  
Major Player ◽  
Ubiquitin Proteasome

The ubiquitin proteasome system (UPS) has emerged from obscurity to be seen as a major player in all regulatory processes in the cell. The concentrations of key proteins in diverse regulatory pathways are controlled by post-translational ubiquitination and degradation by the 26 S proteasome. These regulatory cascades include growth-factor-controlled signal-transduction pathways and multiple points in the cell cycle. The cell cycle is orchestrated by a combination of cyclin-dependent kinases, kinase inhibitors and protein phosphorylation, together with the timely and specific degradation of cyclins and kinase inhibitors at critical points in the cell cycle by the UPS. These processes provide the irreversibility needed for movement of the cycle through gap 1 (G1), DNA synthesis (S), gap 2 (G2) and mitosis (M). The molecular events include cell-size control, DNA replication, DNA repair, chromosomal rearrangements and cell division. It is doubtful whether these events could be achieved without the temporally and spatially regulated combination of protein phosphorylation and ubiquitin-dependent degradation of key cell-cycle regulatory proteins. The oncogenic transformation of cells is a multistep process that can be triggered by mutation of genes for proteins involved in regulatory processes from the cell surface to the nucleus. Since the UPS has critical functions at all these levels of control, it is to be expected that UPS activities will be central to cell transformation and cancer progression.

Immunodetection of androgen and estrogen a,b receptors in human urinary bladder cancer

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 15616-15616
Author(s):  
A. Gómez Pinillos ◽  
G. Bueno Serrano ◽  
S. Sacristán López ◽  
R. García González ◽  
C. Varona Crespo ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Western Blotting ◽  
Cancer Progression ◽  
Transitional Cell ◽  
Urinary Bladder Cancer ◽  
Tumor Grade ◽  
Chi Square ◽  
Primary Tumors ◽  
Female Ratio ◽  
Covalent Modifications

15616 Background: Bladder cancer predominates in males. Loss of androgen receptor (AR) has been associated with invasive and undifferentiated tumors but little is known about the role of hormonal receptors in this type of cancer. Methods: We evaluated samples from 41 nonconsecutive patients treated for bladder transitional cell carcinoma between 2001–2003. Immunohistochemistry was performed using anti-AR and anti-ER a,b antibodies on paraffin-embedded tumors from transurethral resections, nuclear expression was assessed. Western blotting for AR and pAKT were performed in 22 transurethral resections from nonconsecutive bladder cancer patients treated between 2004–2006. Chi-square and U Mann-Witney tests were performed. Results: Immunohistochemistry study: the male/female ratio was 9/1; 61% were noninvasive tumors (pTa, is,1) and 32% were invasive (pT2,3); 22% were G1 and 75% were G2,3. 95% were AR(+), 40% ERa(+) and 94% ERb(+). Significant differences in AR expression were observed between G1 78% and G2,3 tumors 100%, with a high grade tumor prevalence in the AR(+) tumors 81.6% p=0.046. No differences between ERa,b and differentiation were found. G2,3 tumors were predominantly ERb(+). AR was found in 92% noninvasive and in 100% invasive tumors p=0.53. No significant differences were found between invasiveness and ERa,b. Noninvasive tumors showed higher ERa,b expresión (54%, 95% respectively) than invasive tumors (20%, 91% respectively). Western blotting group: 54.5% were primary tumors and 45.5% were recidives. The noninvasive/invasive rate was 59/32%, the G1/G2,3 rate was 4.5/95.5%. 112, 250 and 160 kDa bands were found. AR/pAKT rate was 86.4/100%. 100% of invasive tumors and 86% G2,3 tumors were AR(+). No differences between pAKT, invasiveness, differentiation or AR expression were observed but recidivated tumors had a slight higher median expression level. Conclusions: AR is positively related to tumor grade and might be involved in bladder cancer progression. The profile of AR bands suggest covalent modifications, due to ubiquitin-mediated degradation or to activation mechanisms mediated by SUMO. Nuclear AR and the absence of bands under 90kDa suggest that activated protein is present in bladder cancer. pAKT/AR inhibitors could play a role for bladder cancer therapy. No significant financial relationships to disclose.

scholarly journals The Involvement of Ubiquitination Machinery in Cell Cycle Regulation and Cancer Progression

2021 ◽  
Vol 22 (11) ◽  
pp. 5754
Author(s):  
Tingting Zou ◽  
Zhenghong Lin
Keyword(s):  
Cell Cycle ◽  
Cancer Progression ◽  
Ubiquitin Proteasome System ◽  
High Accumulation ◽  
Cancer Occurrence ◽  
Ubiquitin Proteasome ◽  
Cell Cycle Transition ◽  
Cycle Regulation ◽  
Cellular Components ◽  
Ubiquitination Machinery

The cell cycle is a collection of events by which cellular components such as genetic materials and cytoplasmic components are accurately divided into two daughter cells. The cell-cycle transition is primarily driven by the activation of cyclin-dependent kinases (CDKs), the activities of which are regulated by the ubiquitin-mediated proteolysis of key regulators such as cyclins and CDK inhibitors (CKIs). Thus, the ubiquitin-proteasome system (UPS) plays a pivotal role in the regulation of the cell-cycle process via recognition, interaction, and ubiquitination or deubiquitination of key proteins. The illegitimate degradation of tumor suppressor proteins and oncoproteins or, inversely, abnormally high accumulation results in cell proliferation deregulation, genomic instability, and cancer occurrence. In this review, we demonstrate the diversity and complexity of the UPS machinery regulation of the cell cycle. A profound understanding of the ubiquitination machinery will provide new insights into the regulation of the cell-cycle transition, cancer treatment, and the development of anti-cancer drugs.

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