Flow cytometry as a diagnostic method for colorectal cancer

2012 ◽  
Author(s):  
S. Takeda ◽  
N. Hinata ◽  
H. Kanda ◽  
A. Suzuki ◽  
T. Shioyama ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Flow Cytometry ◽  
Diagnostic Method

Synergistic Effects of 5-Fluorouracil in Combination with Diosmetin in Colorectal Cancer Cells

2021 ◽  
Vol 7 (1) ◽  
pp. 6
Author(s):  
Sareh Kamran ◽  
Ajantha Sinniah ◽  
Mohammed Abdullah Alshawsh
Keyword(s):  
Colorectal Cancer ◽  
Flow Cytometry ◽  
Synergistic Effect ◽  
Cancer Cells ◽  
Synergistic Effects ◽  
Neoadjuvant Treatment ◽  
Software Tool ◽  
New Combination ◽  
Combination Regimen ◽  
Colorectal Cancer Cells

Colorectal cancer (CRC) is among the most commonly occurring cancers. The management of CRC includes laparoscopic surgery, radiotherapy, chemotherapies and neoadjuvant treatment. However, conventional chemotherapies have poor impact on combating CRC and are associated with severe toxic effects and high rates of relapse. Therefore, searching for a new combination regimen is a favorable consideration. The aim of this study was to elucidate the synergistic effect of 5-fluorouracil (5-FU) and diosmetin in an in vitro model on colorectal cancer cells. An MTT assay was conducted on HCT-116 cancer cells and they were treated with a concentration gradient of 5-FU and diosmetin individually and in combination. The combination index (CI) and dose reduction index (DRI) were calculated using CompuSyn software. Isobologram analysis and synergism determination were performed using the Combenefit software tool and the synergy score was calculated using the SynergyFinder 2.0 software tool. The apoptotic features of the cells were determined via an AO/PI double staining assay and an annexin V assay using a fluorescent microscope and the flow cytometry technique, respectively. The findings showed that the DRI of 5-FU was three-fold lower in the combination with a CI value of less than one, which indicates that there was a synergistic effect. The AO/PI microscopic results revealed signs of apoptosis and dead cells after 72 h of treatment. Flow cytometry analysis confirmed that the apoptotic effect of the combination was more prominent compared to 5-FU alone. The findings of this study offer a potential strategy for reducing the cytotoxicity and enhancing the efficacy of 5-FU on colorectal cancer cells through a synergistic study model.

scholarly journals Serum microRNAs: A new diagnostic method for colorectal cancer

2013 ◽  
Vol 1 (4) ◽  
pp. 495-498 ◽  
Author(s):  
YI YANG ◽  
XIAODONG GU ◽  
MINWEI ZHOU ◽  
JIANBIN XIANG ◽  
ZONGYOU CHEN
Keyword(s):  
Colorectal Cancer ◽  
Diagnostic Method ◽  
Serum Micrornas

scholarly journals miR-128 Targets the SIRT1/ROS/DR5 Pathway to Sensitize Colorectal Cancer to TRAIL-Induced Apoptosis

2018 ◽  
Vol 49 (6) ◽  
pp. 2151-2162 ◽  
Author(s):  
Bo Lian ◽  
Dongxiang Yang ◽  
Yanlong Liu ◽  
Gang Shi ◽  
Jibin Li ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Flow Cytometry ◽  
Western Blot ◽  
Cancer Cells ◽  
Cell Lines ◽  
Sirtuin 1 ◽  
Luciferase Reporter ◽  
Pcr Analysis ◽  
Reporter Assays ◽  
Induced Apoptosis

Background/Aims: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is an ideal anti-tumor drug because it exhibits selective cytotoxicity against cancer cells. However, certain cancer cells are resistant to TRAIL, and the potential mechanisms are still unclear. The aim of this study was to reduce the resistance of colorectal cancer (CRC) cells to TRAIL. Methods: Quantitative real-time PCR analysis was performed to detect the expression of microRNA-128 (miR-128) in tissues from patients with CRC and CRC cell lines. MTT assays were used to evaluate the effect of miR-128 on TRAIL-induced cytotoxicity against CRC cell lines. The distribution of death receptor 5 (DR5) and the production of reactive oxygen species (ROS) were detected by flow cytometry analysis. Western blot, flow cytometry, and luciferase reporter assays were performed to evaluate the potential mechanism and pathway of miR-128-promoted apoptosis in TRAIL-treated CRC cells. Results: MiR-128 expression was downregulated in tumor tissues from patients with CRC as well as in CRC cell lines in vitro. The enforced expression of miR-128 sensitized CRC cells to TRAIL-induced cytotoxicity by inducing apoptosis. Mechanistically, bioinformatics, western blot analysis, and luciferase reporter assays showed that miR-128 directly targeted sirtuin 1 (SIRT1) in CRC cells. miR-128 overexpression suppressed SIRT1 expression, which promoted the production of ROS in TRAIL-treated CRC cells. This increase of ROS subsequently induced DR5 expression, and thus increased TRAIL-induced apoptosis in CRC cells. Conclusion: The combination of miR-128 with TRAIL may represent a novel approach for the treatment of CRC.

Protein Profiling and Sizing of Extracellular Vesicles from Colorectal Cancer Patients via Flow Cytometry

ACS Nano ◽  
2018 ◽  
Vol 12 (1) ◽  
pp. 671-680 ◽  
Author(s):  
Ye Tian ◽  
Ling Ma ◽  
Manfei Gong ◽  
Guoqiang Su ◽  
Shaobin Zhu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Flow Cytometry ◽  
Cancer Patients ◽  
Extracellular Vesicles ◽  
Protein Profiling ◽  
Colorectal Cancer Patients

scholarly journals A Novel Method for Colorectal Cancer Screening Based on Circulating Tumor Cells and Machine Learning

Entropy ◽  
2021 ◽  
Vol 23 (10) ◽  
pp. 1248
Author(s):  
Eleana Hatzidaki ◽  
Aggelos Iliopoulos ◽  
Ioannis Papasotiriou
Keyword(s):  
Colorectal Cancer ◽  
Machine Learning ◽  
Flow Cytometry ◽  
Cancer Screening ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Learning Algorithms ◽  
Machine Learning Algorithms ◽  
Svm Classifier ◽  
Significant Information

Colorectal cancer is one of the most common types of cancer, and it can have a high mortality rate if left untreated or undiagnosed. The fact that CRC becomes symptomatic at advanced stages highlights the importance of early screening. The reference screening method for CRC is colonoscopy, an invasive, time-consuming procedure that requires sedation or anesthesia and is recommended from a certain age and above. The aim of this study was to build a machine learning classifier that can distinguish cancer from non-cancer samples. For this, circulating tumor cells were enumerated using flow cytometry. Their numbers were used as a training set for building an optimized SVM classifier that was subsequently used on a blind set. The SVM classifier’s accuracy on the blind samples was found to be 90.0%, sensitivity was 80.0%, specificity was 100.0%, precision was 100.0% and AUC was 0.98. Finally, in order to test the generalizability of our method, we also compared the performances of different classifiers developed by various machine learning models, using over-sampling datasets generated by the SMOTE algorithm. The results showed that SVM achieved the best performances according to the validation accuracy metric. Overall, our results demonstrate that CTCs enumerated by flow cytometry can provide significant information, which can be used in machine learning algorithms to successfully discriminate between healthy and colorectal cancer patients. The clinical significance of this method could be the development of a simple, fast, non-invasive cancer screening tool based on blood CTC enumeration by flow cytometry and machine learning algorithms.

scholarly journals Reovirus sensitizes microsatellite stable colorectal cancer to anti-PD-1 treatment via cross-talk in innate and adaptive immune systems

2021 ◽  
Author(s):  
Titto Augustine ◽  
Peter John ◽  
Tyler Friedman ◽  
Jeeshan Jiffry ◽  
Hillary Guzik ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Flow Cytometry ◽  
Immune Response ◽  
T Cell ◽  
Cell Lines ◽  
Ex Vivo ◽  
In Vivo Models ◽  
Reovirus Infection ◽  
Immune Response Genes ◽  
Anti Tumor Activity

Background: Microsatellite stable (MSS) colorectal cancer (CRC) represents ~85% of all CRCs. These tumors are poorly immunogenic and largely resistant to immunotherapy, necessitating a need to develop new immune enhancing strategies. Oncolytic reovirus has a high propensity to replicate in KRAS mutant tumors which account for ~50% of MSS CRCs. Current study explores the ability of reovirus to potentiate the effect of immune checkpoint inhibition in MSS CRC. Methods: Effectiveness of reovirus infection was quantified through MTT assay for cell viability, and expression of immune-response genes by flow cytometry, RT-qPCR, and microarray. Computational analysis of differentially expressed genes was performed by TAC, DAVID and STRING. Combinatorial approach using anti-PD-1 monoclonal antibody was assessed in ex vivo and in vivo models. Live-cell imaging, tumor volume and survival were measured for quantification of anti-tumor activity. Expression of pattern recognition receptors (PRRs), cell surface and activation markers of immune cells, and PD-1/PD-L1 axis were studied using multi-color flow cytometry, immunoblotting, immunohistochemistry, and immunofluorescence. Results: Reovirus infection exerted growth arrest and expression of immune-response genes in CRCs cell lines in a KRAS-dependent manner. However, microsatellite instability, rather than KRAS status determined immune-repose pathways, functionalities and biological processes post-reovirus infection. Furthermore, reovirus significantly enhanced the anti-tumor activity of anti-human PD-1 [nivolumab] treatment in MSS CRC cell lines ex vivo. Similarly, reovirus increased the activity of anti-mouse PD-1 treatment in the CT26 [MSS, KRASMut], but not the MC38 [MSI, KRASWt] syngeneic mouse model of CRC. Combinatorial treatment has reduced the proliferative index, increased apoptosis and differentially altered PD-L1/PD-1 signaling among CT26 and MC38 tumors. Activation of innate immune system and expression of PRRs and antigen presentation markers were observed under reovirus and anti-PD-1 treatment that additionally reduced immunosuppressive macrophages. This led to an increase in T cell subsets, increase in effector T cell activation, and decrease in exhaustion markers specifically within CT26 microenvironment. Conclusion: The current study systematically evaluates immune characteristics and immune microenvironment of CRC under reovirus/anti-PD-1 combination treatment that proves increased effectiveness among MSS compared to MSI CRCs. This is a promising regimen warranting translation into clinical trials.

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