scholarly journals Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients

Biomolecules ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 16
Author(s):  
Saray Duran-Sanchon ◽  
Elena Vila-Navarro ◽  
Maria Marcuello ◽  
Juan José Lozano ◽  
Jenifer Muñoz ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Research Field ◽  
Circulating Microrna ◽  
Endogenous Control ◽  
Serum Samples ◽  
Liquid Biopsies ◽  
Circulating Mirna ◽  
Polymerase Chain ◽  
Advanced Adenomas ◽  
Colorectal Cancer Patients

Background: Circulating microRNA (miRNA) analysis is a growing research field. However, it usually requires an endogenous control or housekeeping (HK) in order to normalize expression of specific miRNAs throughout different samples. Unfortunately, no adequate HK for circulating miRNA analysis is still known in the colorectal cancer (CRC) context whereas several have been suggested. Hence, our aims were to validate the previously suggested miR-1228-3p as HK for CRC studies, to compare its suitability with the widely used miR-16-5p, and to evaluate the influence of hemolysis on both miRNAs. Methods: We analyzed by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) the expression of miR-1228-3p, miR-16-5p and the spike-in cel-miR-39 in a set of 297 plasmas (92 CRC, 101 advanced adenomas -AA-, and 100 controls) and 213 serum samples (59 CRC, 74 AA and 80 controls). We also analyzed both miRNAs depending on the hemolysis degree in 7 plasmas and 31 serums. Results: Levels of miR-1228-3p and miR-16-5p did not show significant differences between groups although miR-16-5p exhibited more variability in plasma and serum samples. Importantly, the combination of cel-miR-39 and miR-1228-3p was the most stable one. Moreover, we observed that miR-16-5p was significantly influenced by hemolysis in contrast with miR-1228-3p that exhibited no correlation with this confounding factor in both biofluids. Conclusion: MiR-1228-3p has been validated as an adequate endogenous control for circulating miRNA analysis in CRC and AA liquid biopsies.

scholarly journals Correlation between IL-10 as Cancer Biomarker and Demographic Characteristics of Colorectal Cancer Patients

2021 ◽  
pp. 1-10
Author(s):  
Rahin Sh Hamad ◽  
Bushra H. Shnawa ◽  
Shereen J. Al-Ali
Keyword(s):  
Colorectal Cancer ◽  
Pearson Correlation ◽  
Serum Levels ◽  
Interleukin 10 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
High Morbidity ◽  
Serum Samples ◽  
Cancer Pathogenesis ◽  
Colorectal Cancer Patients

Colorectal cancer (CRC) is classified as one of the most prevalent cancer types worldwide, with high morbidity and mortality rates. Patients of CRC have been shown to express a detectable cytokine in serum which contributes to cancer pathogenesis. Therefore, the serum interleukin 10 (IL-10) level in CRC patients was investigated in this study. Patients' medical records with CRC admitted to the Rizgary and Nanakali hospitals, Erbil, Iraq was analyzed as the study group compared to the healthy volunteers' control group. Seventy-one serum samples were collected, thirty-one from diagnosed CRC patients and forty from healthy controls. The concentrations of IL-10 in the sera were assessed by enzyme-linked immunosorbent assay (ELISA). The present finding showed that IL-10 Was significantly elevated in CRC patients' sera compared to the control group, suggesting confirmation of its usefulness for detecting CRC patients' prognosis. A non-significant Pearson correlation was detected between IL-10 serum levels and the CRC group's age, gender, and body mass index. Herein is the first study on the evaluation of IL-10 levels in CRC patients in Kurdistan, Iraq.

scholarly journals Facile profiling of molecular heterogeneity by microfluidic digital melt

2018 ◽  
Vol 4 (9) ◽  
pp. eaat6459 ◽  
Author(s):  
Christine M. O’Keefe ◽  
Thomas R. Pisanic ◽  
Helena Zec ◽  
Michael J. Overman ◽  
James G. Herman ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Tumor Suppressor ◽  
Tumor Suppressor Gene ◽  
Single Molecule ◽  
Rapid Assessment ◽  
Suppressor Gene ◽  
Molecular Heterogeneity ◽  
Liquid Biopsies ◽  
Digital Microfluidic ◽  
Colorectal Cancer Patients

This work presents a digital microfluidic platform called HYPER-Melt (high-density profiling and enumeration by melt) for highly parallelized copy-by-copy DNA molecular profiling. HYPER-Melt provides a facile means of detecting and assessing sequence variations of thousands of individual DNA molecules through digitization in a nanowell microchip array, allowing amplification and interrogation of individual template molecules by detecting HRM fluorescence changes due to sequence-dependent denaturation. As a model application, HYPER-Melt is used here for the detection and assessment of intermolecular heterogeneity of DNA methylation within the promoters of classical tumor suppressor genes. The capabilities of this platform are validated through serial dilutions of mixed epialleles, with demonstrated detection limits as low as 1 methylated variant in 2 million unmethylated templates (0.00005%) of a classic tumor suppressor gene,CDKN2A(p14ARF). The clinical potential of the platform is demonstrated using a digital assay forNDRG4, a tumor suppressor gene that is commonly methylated in colorectal cancer, in liquid biopsies of healthy and colorectal cancer patients. Overall, the platform provides the depth of information, simplicity of use, and single-molecule sensitivity necessary for rapid assessment of intermolecular variation contributing to genetic and epigenetic heterogeneity for challenging applications in embryogenesis, carcinogenesis, and rare biomarker detection.

scholarly journals Value of Serum NEUROG1 Methylation for the Detection of Advanced Adenomas and Colorectal Cancer

Diagnostics ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 437
Author(s):  
Olalla Otero-Estévez ◽  
María Gallardo-Gomez ◽  
María Páez de la Cadena ◽  
Francisco Javier Rodríguez-Berrocal ◽  
Joaquín Cubiella ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cpg Island ◽  
Family Risk ◽  
Liquid Biopsies ◽  
Age And Gender ◽  
Screening Programs ◽  
Aberrant Dna Methylation ◽  
And Gender ◽  
Asymptomatic Individuals ◽  
Advanced Adenomas

Aberrant DNA methylation detected in liquid biopsies is a promising approach for colorectal cancer (CRC) detection, including premalignant advanced adenomas (AA). We evaluated the diagnostic capability of serum NEUROG1 methylation for the detection of AA and CRC. A CpG island in NEUROG1 promoter was assessed by bisulfite pyrosequencing in a case-control cohort to select optimal CpGs. Selected sites were evaluated through a nested methylation-specific qPCR custom assay in a screening cohort of 504 asymptomatic family-risk individuals. Individuals with no colorectal findings and benign pathologies showed low serum NEUROG1 methylation, similar to non-advanced adenomas. Contrarily, individuals bearing AA or CRC (advanced neoplasia—AN), exhibited increased NEUROG1 methylation. Using >1.3518% as NEUROG1 cut-off (90.60% specificity), 33.33% of AN and 32.08% of AA were identified, detecting 50% CRC cases. Nonetheless, the combination of NEUROG1 with fecal immunochemical test (FIT), together with age and gender through a multivariate logistic regression resulted in an AUC = 0.810 for AN, and 0.796 for AA, detecting all cancer cases and 35–47% AA (specificity 98–95%). The combination of NEUROG1 methylation with FIT, age and gender demonstrated a convenient performance for the detection of CRC and AA, providing a valuable tool for CRC screening programs in asymptomatic individuals.

scholarly journals Untargeted Metabolomics and Polyamine Profiling in Serum before and after Surgery in Colorectal Cancer Patients

Metabolites ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 487
Author(s):  
Yu Ra Lee ◽  
Ki-Yong An ◽  
Justin Jeon ◽  
Nam Kyu Kim ◽  
Ji Won Lee ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Metabolic Pathways ◽  
Sphingolipid Metabolism ◽  
Proline Metabolism ◽  
Liquid Chromatography Mass Spectrometry ◽  
Targeted Metabolomics ◽  
Serum Samples ◽  
Physiological Alterations ◽  
Before And After ◽  
Colorectal Cancer Patients

Colorectal cancer is one of the most prevalent cancers in Korea and globally. In this study, we aimed to characterize the differential serum metabolomic profiles between pre-operative and post-operative patients with colorectal cancer. To investigate the significant metabolites and metabolic pathways associated with colorectal cancer, we analyzed serum samples from 68 patients (aged 20–71, mean 57.57 years). Untargeted and targeted metabolomics profiling in patients with colorectal cancer were performed using liquid chromatography-mass spectrometry. Untargeted analysis identified differences in sphingolipid metabolism, steroid biosynthesis, and arginine and proline metabolism in pre- and post-operative patients with colorectal cancer. We then performed quantitative target profiling of polyamines, synthesized from arginine and proline metabolism, to identify potential polyamines that may serve as effective biomarkers for colorectal cancer. Results indicate a significantly reduced serum concentration of putrescine in post-operative patients compared to pre-operative patients. Our metabolomics approach provided insights into the physiological alterations in patients with colorectal cancer after surgery.

Evaluation of serum CXCL5 as a serum marker for prognosis of colorectal cancer patients.

2011 ◽  
Vol 29 (4_suppl) ◽  
pp. 442-442 ◽  
Author(s):  
M. Kawamura ◽  
Y. Toiyama ◽  
K. Tanaka ◽  
H. Yasuda ◽  
H. Fujikawa ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Overall Survival ◽  
Liver Metastasis ◽  
Cancer Patients ◽  
Normal Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Serum Samples ◽  
Preoperative Serum ◽  
Colorectal Cancer Patients

442 Background: CXCL5 is known as CXC chemokine which promotes angiogenesis related to cancer. However, the function of serum level of CXCL5 (sCXCL5) has not been fully studied in colorectal cancer. The purpose of this study was to evaluate the relationships between preoperative sCXCL5 and clinicopathological features and prognosis in colorectal cancer. Methods: This was a single-institution, retrospective study. Preoperative serum samples of 250 colorectal cancer patients (between 1998 and 2007, median age: 65.3 years, male 159/female 91) were available for the study, and 33 normal serum was examined and used as a control. sCXCL5 level was assayed using a commercially available enzyme-linked immunosorbent assay kit, and analyzed statistically. Results: Mean level of sCXCL5 was significantly higher in colorectal cancer patients than in control group (p=0.013). Patients with liver metastases had significantly higher sCXCL5 level than those without metastases (p=0.0086), and in logistic analysis, sCXCL5 was an independent marker for predicting liver metastasis (p=0.040). Overall survival of patients with elevated sCXCL5 level was significantly worse than those with lower sCXCL5 (p=0.0006). Conclusions: Preoperative sCXCL5 level was increased in colorectal cancer patients compared to in healthy volunteer and elevated sCXCL5 was correlated with liver metastasis and poor prognosis for overall survival in colorectal cancer patients. Elevated sCXCL5 has been proposed as a useful predictive marker for liver metastasis and overall survival in colorectal cancer. No significant financial relationships to disclose.

scholarly journals miR-124 Functions as A Marker in Diagnosing Colorectal Cancer

2020 ◽  
Author(s):  
Yang Yan ◽  
Xiaohui Du ◽  
Shaoyou Xia ◽  
Songyan Li ◽  
Da Teng ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Roc Curve ◽  
Operating Characteristic ◽  
Chain Reaction ◽  
Serum Samples ◽  
Optimal Cutoff ◽  
Clinicopathologic Factors ◽  
Healthy Control ◽  
Optimal Cutoff Point ◽  
Polymerase Chain

Abstract Background: MicroRNA-124 (miR-124) regulates cell growth, angiogenesis, invasion, and apoptosis of many human tumors. Our objective was to evaluate the levels of miR-124 in serum samples of colorectal cancer (CRC) patients to explore whether miR-124 in serum can be used as a biomarker for the detection of CRC.Methods: Serum miR-124 level was measured in 85 patients with CRC and 60 healthy control subjects using reverse transcription quantitative real-time polymerase chain reaction (qRT-PCR). The correlations between miR-124 levels and clinicopathologic factors were analyzed. Diagnostic performance of serum miR-124 level was calculated by using the receiver operating characteristic (ROC) curve.Results: The expression level of miR-124 in serum samples was significantly lower in CRC patients than in healthy controls (P<0.001). A positive correlation between miR-124 level and tumor size (P=0.010), invasion depth (P=0.021), lymph node metastasis (P=0.015), and TNM stage (P=0.004) was observed. The serum miR-124 yielded an AUC (areas under the ROC curve) of 0.832 with a sensitivity of 80.0% and a specificity of 81.7%, and the optimal cutoff point of miR-124 was 1.61.Conclusions: Serum miR-124 may have a potential as a novel biomarker for the detection of CRC.

Prognostic value of circulating cytokines in colorectal cancer: A prospective study in sixty colorectal cancer patients in Tunisia.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e15130-e15130
Author(s):  
Jihene Braham Ayari ◽  
Rania Guesmi ◽  
Mehdi Balti ◽  
Mouna Ben Azaiz ◽  
Aref Zribi ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cancer Patients ◽  
Solid Phase ◽  
Young Patients ◽  
Serum Samples ◽  
Metastatic Setting ◽  
The Mean ◽  
High Level ◽  
Colorectal Cancer Patients ◽  
Circulating Cytokines

e15130 Background: Colorectal cancer is the third most common malignancy and fourth most common cause of cancer mortality worldwide. It is responsible for more than 600,000 deaths annually, and incidence rates are increasing in most of the developing countries. Pathophysiology implicates pro-inflammatory conditions that promote the tumor malignant progression, invasion, and metastasis. The aim of this study is to measure the level of circulating cytokines (IL1b, IL6, IL8, IL10, IL22, IL23 and TNFα) in sixty colorectal cancer patients in Tunisia and to evaluate their implication as prognostic factors. Methods: Serum samples were collected prospectively from a cohort of sixty colorectal cancer patients in Tunisia. Levels of circulating inflammatory cytokines, TNF-α, IL1b, IL6 and IL8 were measured using the technique of a solid-phase, two-site chemo-luminescent enzyme immune-metric assay (Immulite 1000, Simens, USA). Serum levels of IL10 were measured by enzyme-linked immunosorbent assays (ELISA) sandwich method. Results: The mean age of patients is 58 years (24–82 years), Thirty-sex among them were m and 24 women with sex ratio of 1.5. Twenty-five patients were at metastatic setting, and hepatic metastasis was found in 25% of cases. The mean level of cytokines Il6, IL10, TNFα, IL8 and IL1b were respectively 12.26 +/- 18.7 pg/ ml (min 2, max 117pg/ ml), 0.93 +/- 5.23 pg/ ml (min 0, max 39.35 pg/ml), 8.31 +/- 4.99 pg/ ml (min 4, max 27.20 pg/ ml), 61.9 +/- 159.71 pg/ml (min 5, max 1173 pg/ ml) and 1.13 +/- 3.34 pg/ ml (min 5, max 15.7pg/ml. We found a significant correlation between a high level of IL8 and metastatic disease (p=0.001), especially in mutant RAS cases (p=0.001). We found also a significant correlation between high level of IL1b and lymphovascular invasion (p=0.013) and young patients (p=0.01). On the other hand, there was significant correlation between IL8 and IL6 (r = 0.560, p = 0.00001); IL8 and TNFα (r = 0.404, p = 0.001); and IL10 with IL1b (r = 0.297, p = 0.021). Conclusions: Our results highlight the role of circulating IL8, TNFα, IL1b and IL10 as potential prognostic biomarkers in colorectal cancer patients. These cytokines could contribute to tumor growth and progression, namely for IL-8 level that was significantly correlated with poor prognosis and advanced stages. This correlation needs to be evaluated in large prospective trials and suggests a rational for the development and use of cytokine blockade in treatment of colorectal cancer patients.

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