Abstract Objectives TIMPs control the activity of MMPs, one of the key molecules for tumor invasion and metastasis. The aim of this study was to assess the usefulness of MMP-2 and MMP-9 in relation ...to their inhibitor (TIMP2) as noninvasive diagnostic tests for bilharzial bladder cancer. Material and methods Voided urine samples were provided from 244 subjects (154 bladder cancer 136 bilharzial; 60 benign urologic disorders; 30 healthy volunteers). Urine sediment was used for cytology, and the supernatant for estimation of MMPs and TIMP-2 by ELISA and gelatin zymography. Results The best cut-off values for the investigated markers were determined by ROC curve. Positivity rates and median levels for MMP-2, MMP-9, TIMP-2, MMP-2/TIMP-2, and MMP-9/TIMP-2 showed significant difference among the three investigated groups ( p < 0.001). MMP-9 and MMP-2/TIMP-2 were related to pathologic type, MMP-2/TIMP-2 was inversely related to the grade, and MMP-9/TIMP-2 was related to bilharziasis ( p < 0.05). MMP zymography results were comparable to those from ELISA. Conclusion The sensitivity and specificity of MMP zymography, MMP-9/TIMP-2 ratio, and MMP-2/TIMP2 ratio were superior among all investigated parameters; furthermore, combined testing of cytology with them improves the sensitivity even in superficial and low-grade tumors.
Early screening for bladder cancer (BC) holds the key to combat and control the increasing global burden of BC mortality. We presented a simple approach to characterize, analyze, and validate a panel ...of biomarkers in BC and their relationship to bilharziasis. We investigated voided urine and blood samples from patients with bladder cancer (
n
= 94), benign bladder lesions (
n
= 60), and age-matched normal controls (
n
= 56). This study was divided into the following phases. (1) We analyzed the expression of urinary Hyaluronoglucosaminidase 1 (HYAL1) protein in BC and control samples by zymography. (2) We performed bioinformatics analysis to retrieve a set of epigenetic regulators of HYAL1. (3) This set of three selected genes long non-coding RNA-urothelial cancer associated 1(
lncRNA
-UCA1), microRNA-210, and microRNA-96 was then analyzed in the same urine samples used in phase I by quantitative real-time PCR. (4) A high reproducibility of gene selection results was also determined from statistical validation. The urinary expression of HYAL1 protein and its epigenetic regulators were higher in BC patients (
P
< .001). The receiver-operating characteristic curve analyses demonstrated that each one had good sensitivity and specificity for distinguishing BC patients from non-BC ones (HYAL1, 89.4 and 91.2 %; miR-210, 76.6 and 93 %;
miR
-96, 76.6 and 89.4 %; and
lncRNA
-UCA1, 91.5 and 96.5 %). There was a significant positive correlation between HYAL1 and the selected epigenetic biomarkers. The performance of this urine biomarker panel reached 100 % sensitivity and 89.5 % specificity for bladder cancer diagnosis.
Matrix metalloproteinases (MMPs), in particularly gelatinases (MMP-2 and MMP-9) were reported as urinary markers of bladder cancer. In this work, we developed a simple colorimetric gold nanoparticle ...(AuNP) assay for rapid and sensitive detection of urinary total gelatinase activity based on the surface plasmon resonance (SPR) property of AuNPs. Gelatin-modified AuNPs were stably suspended in solution even upon addition of an aggregation inducer as 6-mercaptohexan-1-ol (6-MCH). Gelatinases digest gelatin capping. Subsequently, addition of 6-MCH leads to AuNPs aggregation with red to blue color shift. In a pilot study, results of the developed AuNP assay were consistent with zymography for qualitative detection of urinary total gelatinase activity. The sensitivity and specificity of both assays were 80% and 90.9% respectively. The absorption ratios, A625/A530 of the reacted AuNP solutions were used to quantify the total gelatinase concentration. The best cut off value was 0.01895ng/μg protein, at which the sensitivity was 87.5% and the specificity was 86.4%. The developed AuNP assay is simple, low-cost and can aid non-invasive diagnosis of bladder cancer.
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•A simple gelatin-modified AuNP biosensor was developed for rapid and sensitive detection of urinary total gelatinase activity.•The developed AuNP assay results were consistent with zymography for qualitative detection of urinary total gelatinase activity.•The sensitivity of both AuNP assay and zymography was 80%, while the specificity for both assays was 90.9%.•The developed gelatin-modified AuNP platform can aid non-invasive diagnosis of bladder cancer.
We investigated the action of caffeic acid in regulating miR-636 expression level in kidney of streptozotocin-induced diabetic rats. Streptozotocin-induced diabetic rats were orally treated with ...caffeic acid at 40 mg/kg/day for 8 weeks. At the end of the treatment, body and kidney weight and blood glucose levels were determined, blood, urine, and kidneys were collected for biochemical and histological examination. Expression levels of miR-636 were determined in liver by qRT-PCR. Induction of diabetic nephropathy by streptozotocin was evidenced by displayed elevated levels of serum creatinine, blood urea nitrogen, microalbuminuria and urinary albumin/creatinine ratio in addition to renal hypotrophy. Caffeic acid (CA) can ameliorate renal damage and significantly decreased the fasting blood glucose, cholesterol and triglyceride in diabetic rats. CA treatment improved histological architecture in the diabetic kidney. CA significantly down regulate miR-636 expression level in the kidney of diabetic rats in comparison to healthy group. Overall, caffeic acid down regulates miR-636 expression level which is involved in development of diabetic nephropathy and might therefore be potential attractive therapeutic agent to pursue in DN.
We assessed the differential expression of a urinary panel of microRNAs (miRs) in terms of potential application as diagnostic markers of bladder cancer (BC) and relationship to bilharziasis. We ...investigated voided urine samples and blood from patients with BC (n = 188), benign bladder lesions (n = 88), and age-matched controls (n = 92). Five miRs (miR-210, miR-10b, miR-29c, miR-221, and miR-23a) were selected from previous microarray signature profiling (released by miR2Disease). Afterward, they were validated using polymerase chain reaction array. The expression levels of miR-210, miR-10b, and miR-29c in the urine samples were significantly higher in BC ( P < 0.001). The receiver-operating characteristic curve analyses demonstrated that each miR had good sensitivity and specificity for distinguishing patients with BC from patients without BC (miR-210, 71.3% and 91.1%; miR-10b, 80.9% and 91.1%; and miR-183, 71.3% and 88.9%). On combining the 3 miR detection data with the urinary cytology, the results sensitivity increased to 95.2%. Relative quantity mean rank of the miR-29c was significantly higher in the bilharzial-positive patients compared with bilharzial-negative patients. To conclude, urine miR-210, miR-10b, and miR-29c are promising tumor markers for BC: bilharzial and nonbilharzial.
Colorectal cancer (CRC) is characterized by high heterogeneity and a complex microenvironment that leads to high inter-patient variability. Personalized management of CRC could address this. ...Accumulating data highlights the interaction between the CRC microenvironment and the immune system through different cells and receptors with a focus on the toll-like receptors (TLRs). Multiple studies identified a bidirectional role played by TLRs in CRC with involvement in both carcinogenesis and therapy.
A study to highlight the interaction between TLRs and CRC microenvironment on different molecular levels was undertaken, addressing TLR gene polymorphism, TLR genetic and epigenetic deregulation and TLR ligand binders. In addition, the use of these TLRs and their interaction with CRC microenvironment were evaluated to identify novel CRC therapeutics.
Previous literature has shown that TLRs are incriminated in CRC pathogenesis and thus research effort was directed to make use of these TLRs in drawing new therapeutic patterns for CRC. However, to date, these immune-therapeutic patterns of CRC have shown limited success in reducing tumor burden. This highlights the need for more studies that would better illustrate the interaction between TLRs and CRC microenvironment and the impact of TLR modifications to yield more efficient and precise CRC therapeutics.
The aim of this study is to identify micro-ribonucleic acid (microRNA) and its target, in addition to their relationship to the outcome in breast cancer (BC). To achieve this aim, we investigated ...microRNA-10b (miR-10b) and minichromosome maintenance complex component 5 (
MCM5
mRNA) expression in 230 breast tissue samples by real-time PCR and semiquantitative conventional RT-PCR, respectively. Relapse-free survival (RFS) associated with miRNA-10b and
MCM5
mRNA were tested by Kaplan–Meier survival analysis. The impact of miRNA-10b and
MCM5
mRNA expression on the survival was evaluated by Cox proportional hazard regression model. The expression of miRNA-10b and
MCM5
mRNA was positive in 86.4 and 79.7 % breast cancer patients, respectively. The overall concordance rate between miRNA-10b and
MCM5
RNA was 90.4 %. The median follow-up period was 50 months. The survival analysis showed that high levels of both miR-10b and
MCM5
were associated with short relapse free survival of BC. We identified
MCM5
mRNA expression changes consistent with the miRNA-10b target regulation. Thus, we could consider miRNA-10b and
MCM5
mRNA as prognostic markers and potential therapeutic targets in breast cancer to be applied to other patient data sets.
To develop a gold nanoparticle (AuNP) assay for direct detection of unamplified HURP RNA in urine.
HURP RNA was extracted from urine samples (50 bladder carcinoma patients, 25 benign bladder lesions, ...and 25 controls) and further purified using magnetic nanoparticles (MNPs), functionalized with HURP RNA-specific oligonucleotides, and then detected by RT-PCR or gold nanoparticles.
The developed HURP RNA AuNP assay has a sensitivity and a specificity of 88.5% and 94%, respectively, and a detection limit of 2.4nmol/L. The concordance between the HURP AuNP assay with RT-PCR after RNA purification using functionalized MNPs was 97%.
The developed colorimetric HURP RNA AuNP assay is sensitive, simple, and can aid noninvasive diagnosis of bladder cancer.
•A simple, sensitive, rapid AuNP assay was developed for detection of HURP RNA.•The assay employs magnetic and AuNPs for purification and detection of HURP RNA.•The assay has 88.5%, sensitivity, 94% specificity, and 2.4nmol/L detection limit.•The concordance with RT-PCR after RNA purification using magnetic NPs was 97%.•The new platform could aid noninvasive diagnosis of bladder cancer.
Purpose Urinary tumor markers that help in the early detection of bladder cancer promise a significant improvement in sensitivity, specificity and convenience over conventional, invasive diagnostic ...tests. We assessed the diagnostic efficacy of hyaluronidase (HYAL1) and survivin for early bladder cancer detection. Materials and Methods The study included 166 patients diagnosed with bladder carcinoma, 112 with benign bladder lesions and 100 healthy volunteers who served as controls. All underwent serological assessment of schistosomiasis antibody, urine cytology, and hyaluronidase (HYAL1) and survivin RNA estimation by qualitative and semiquantitative reverse transcriptase-polymerase chain reaction in urothelial cells from voided urine. Results Positivity rates of HYAL1 RNA and survivin RNA on qualitative reverse transcriptase-polymerase chain reaction were significantly different among the 3 groups. Mean rank using semiquantitative method was increased in the malignant vs the other groups. The best cutoff for HYAL1 and survivin RNA was 0.25 each. Using these cutoffs HYAL1 and survivin RNA sensitivity was 91% and 75%, respectively, with absolute specificity. HYAL1 RNA detected all patients with stages 0 and I bladder cancer (p <0.037). Urine cytology sensitivity improved when combined with hyaluronidase or survivin RNA on semiquantitative reverse transcriptase-polymerase chain reaction. Conclusions The detection of urinary HYAL1 and survivin RNA is a promising noninvasive test for bladder cancer early detection. HYAL1 RNA was more sensitive and specific than urine cytology. Semiquantitative reverse transcriptase-polymerase chain reaction is favored for its high sensitivity and specificity.
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