scholarly journals Comparative analysis of the bioenergetics of human adenocarcinoma Caco-2 cell line and postoperative tissue samples from colorectal cancer patients

2018 ◽  
Vol 96 (6) ◽  
pp. 808-817 ◽  
Author(s):  
Lyudmila Ounpuu ◽  
Laura Truu ◽  
Igor Shevchuk ◽  
Vladimir Chekulayev ◽  
Aleksandr Klepinin ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Line ◽  
Respiratory Chain ◽  
Growth Conditions ◽  
Control Analysis ◽  
Respiratory Chain Complexes ◽  
Colon Tissue ◽  
Tissue Samples

The aim of this work was to explore the key bioenergetic properties for mitochondrial respiration in the widely-used Caco-2 cell line and in human colorectal cancer (HCC) postoperational tissue samples. Oxygraphy and metabolic control analysis (MCA) were applied to estimate the function of oxidative phosphorylation in cultured Caco-2 cells and HCC tissue samples. The mitochondria of Caco-2 cells and HCC tissues displayed larger functional activity of respiratory complex (C)II compared with CI, whereas in normal colon tissue an inverse pattern in the ratio of CI to CII activity was observed. MCA showed that the respiration in Caco-2 and HCC tissue cells is regulated by different parts of electron transport chain. In HCC tissues, this control is performed essentially at the level of respiratory chain complexes I–IV, whereas in Caco-2 cells at the level of CIV (cytochrome c oxidase) and the ATP synthasome. The differences we found in the regulation of respiratory chain activity and glycose index could represent an adaptive response to distinct growth conditions; this highlights the importance of proper validation of results obtained from in-vitro models before their extrapolation to the more complex in-vivo systems.

scholarly journals Upregulation of SNTB1 Correlates with Poor Prognosis and Promotes Cell Growth in Colorectal Cancer

2021 ◽  
Author(s):  
Liya Liu ◽  
Youqin Chen ◽  
Xiaoying Lin ◽  
Meizhu Wu ◽  
Jiapeng Li ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Cycle ◽  
Colony Formation ◽  
Malignant Tumors ◽  
Cdna Array ◽  
Cell Counting ◽  
Nude Mouse Model ◽  
Tissue Samples

Abstract Background: Colorectal cancer (CRC) is one of the most highly malignant tumors and has a complicated pathogenesis. A preliminary study identified syntrophin beta 1 (SNTB1) as a potential oncogene in CRC. However, the clinical significance, biological function, and underlying mechanisms of SNTB1 in CRC are unknown. Thus, the present study aimed to investigate the function of SNTB1 in CRC.Methods: The expression profile of SNTB1 in CRC samples was evaluated by database analysis, cDNA array, tissue microarray, Quantitative real-time PCR (qPCR), and immunohistochemistry. SNTB1 expression in human CRC cells was silenced using short hairpin RNAs and its mRNA and protein levels were assessed by qPCR and western blotting, respectively. Cell proliferation, colony formation, cell cycle and apoptosis were determined by the cell counting, colony formation, and flow cytometry assays, respectively. A xenograft nude mouse model of CRC was established for validating the roles of SNTB1 in vivo. Immunohistochemistry was used to score the expression of SNTB1 in tissue samples. The isobaric tags for relative and absolute quantification (iTRAQ) was used to analyze the differentially expressed proteins after knockdown of SNTB1 in CRC cells.Results: SNTB1 expression was increased in CRC tissue compared with adjacent noncancerous tissues and the increased expression was associated with shorter overall survival of CRC patients. Silencing of SNTB1 suppressed cell viability and survival, induced cell cycle arrest and apoptosis in vitro, and inhibited the growth of CRC cells in vivo. Further elucidation of the regulation of STNB on CRC growth by iTRAQ analysis identified 210 up-regulated and 55 down-regulated proteins in CRC cells after SNTB knockdown. A PPI network analysis identified protein kinase N2 (PKN2) as a hub protein and was up-regulated in CRC cells after SNTB1 knockdown. Western-blot analysis further confirmed that SNTB1 knockdown significantly up-regulated PKN2 protein expression in CRC cells and decreased the phosphorylation of both ERK1/2 and AKT. Conclusion: These findings indicate that SNTB1 is overexpressed in CRC. Elevated SNTB1 levels are correlated with shorter patient survival. Importantly, SNTB1 promoted tumor growth and progression of CRC, possibly by reducing the expression of PKN2 and activating the ERK and AKT signaling pathway. Our study highlights the potential of SNTB1 as a new prognostic predictor and therapeutic target for CRC.

scholarly journals UM-Chor1: establishment and characterization of the first validated clival chordoma cell line

2018 ◽  
Vol 128 (3) ◽  
pp. 701-709 ◽  
Author(s):  
John Henry Owen ◽  
Christine M. Komarck ◽  
Anthony C. Wang ◽  
Waleed M. Abuzeid ◽  
Richard F. Keep ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Malignant Tumors ◽  
Axial Skeleton ◽  
Growth Conditions ◽  
Laboratory Research ◽  
Serum Free ◽  
Clival Chordoma

OBJECTIVEChordomas are rare malignant tumors thought to arise from remnants of the notochord. They can be located anywhere along the axial skeleton but are most commonly found in the clival and sacrococcygeal regions, where the notochord regresses during fetal development. Chordomas are resistant to many current therapies, leaving surgery as the primary method of treatment. Cancer cell lines have been useful for developing new cancer treatments in a laboratory setting that can then be transferred to the clinic, but there are only 4 validated chordoma cell lines available. The objective of this work was to establish chordoma cell lines from surgical tissue in order to expand the library of lines available for laboratory research.METHODSChordoma tissue from the clivus was processed and sorted by flow cytometry to obtain an isolated population of chordoma cells. These cells were grown in culture and expanded until enough doublings to consider the line established. Identification of a chordoma cell line was made with known markers for chordoma, and the line was observed for ALDH (aldehyde dehydrogenase) subpopulations and tested in serum-free growth conditions as well as in vivo.RESULTSA fifth chordoma cell line, UM-Chor1, was successfully established. This is the first chordoma cell line originating from the clivus. Validation was confirmed by phenotype and positivity for the chordoma markers CD24 and brachyury. The authors also attempted to identify an ALDHhigh cell population in UM-Chor1, UCH1, and UCH2 but did not detect a distinct population. UM-Chor1 cells were able to form spheroids in serum-free culture, were successfully transduced with luciferase, and could be injected parasacrally and grown in NOD/SCID mice.CONCLUSIONSThe availability of this novel clival chordoma cell line for in vitro and in vivo research provides an opportunity for developments in treatment against the disease.

scholarly journals Ubiquitin-Like Protein UBD Promotes Cell Proliferation in Colorectal Cancer by Facilitating p53 Degradation

2021 ◽  
Vol 11 ◽  
Author(s):  
Hongbin Su ◽  
Mengdi Qin ◽  
Qiang Liu ◽  
Bo Jin ◽  
Xianjun Shi ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Western Blotting ◽  
Xenograft Model ◽  
Proteasome Inhibition ◽  
Tissue Samples ◽  
Functional Changes ◽  
Tumor Tissues ◽  
Expression Plasmids

PurposeUbiquitin D (UBD) is a member of the ubiquitin-like modifier (UBL) family and is highly expressed in a variety of cancers including colorectal cancer (CRC). However, the mechanisms of its regulatory roles in CRC are largely elusive. In this study, we revealed the effect of UBD on the proliferation of CRC.MethodsThe expression of UBD in clinical tissue samples of CRC and seven CRC cell lines was detected using qRT-PCR, immunohistochemistry (IHC) and Western blotting. CCK-8, colony formation, EdU and flow cytometry assays were used to detect the functional changes of CRC cells transfected with UBD stable expression plasmids in vitro. A xenograft model was constructed to assess the effect of UBD on the growth of CRC cells in vivo. The connection between UBD and p53 was analyzed using Western blotting, immunoprecipitation, proteasome inhibition assay and Cycloheximide (CHX) chase assay.ResultsUBD was overexpressed in CRC tumor tissues compared with nontumor tissues, and its overexpression was positively associated with the tumor size and TNM stage of CRC patients. Functionally, UBD significantly accelerated CRC cell viability and proliferation in vitro and promoted tumorigenesis in vivo. Mechanistically, UBD interacted with p53 in CRC cells, downregulated the expression of p53 by regulating its degradation, shortened the p53 half-life, thereby further affecting the decrease in p21 and the increase in Cyclin D1, Cyclin E, CDK2, CDK4 and CDK6. Moreover, in vivo experiments showed that UBD-induced tumor growth in nude mice was dependent on a decrease in p53.ConclusionsOur study proved that UBD mediates the degradation of p53, thereby facilitating the growth of CRC cells and ultimately promoting the progression of CRC. Therefore, UBD may be a potential therapeutic target and a promising prognostic biomarker for CRC.

scholarly journals The SLC34A2-ROS-HIF-1-induced up-regulation of EZH2 expression promotes proliferation and chemo-resistance to apoptosis in colorectal cancer

2019 ◽  
Vol 39 (5) ◽  
Author(s):  
Xu Li ◽  
Junjie Xing ◽  
Hantao Wang ◽  
Enda Yu
Keyword(s):  
Colorectal Cancer ◽  
Cell Line ◽  
Molecular Mechanisms ◽  
Hypoxia Inducible Factor ◽  
Site Directed Mutagenesis ◽  
Epithelial Cell Line ◽  
Ros Generation ◽  
Ezh2 Expression

AbstractGrowing evidence has uncovered that SLC34A2 plays an evident role in the progression in several types of tumors. However, the biological function and underlying molecular mechanisms of SLC34A2 remain largely unknown. Here, we indicated that SLC34A2 expression was markedly increased in SW480 and HT29 cell line cells compared with that in normal colorectal epithelial cell line cells. Array analysis displayed that the expression of enhancer of zeste 2 (EZH2) decreased considerably when SLC34A2 was knocked down. We demonstrated that SLC34A2 induced EZH2 expression and activated its promoter activity. Serial 5′ deletion and site-directed mutagenesis revealed that the induction of EZH2 expression by SLC34A2 was dependent upon the hypoxia-inducible factor 1 (HIF-1)-2 binding site directly within EZH2 promoter. Moreover, HIF-1 activation was proved essential for SLC34A2-induced EZH2 expression. Reactive oxygen species (ROS) generation contributed to the stabilization of HIF-1α by leading to the binding of HIF-1α to the EZH2 promoter, which resulted in increased EZH2 expression. Additionally, we showed that the inhibition of both HIF-1α expression and ROS generation by YC-1 or BHA, respectively, decreased SLC34A2-induced EZH2 overexpression. Significantly, SLC34A2-induced EZH2 overexpression promoted the proliferation and chemo-resistance to apoptosis in colorectal cancer (CRC) cells in vitro and in vivo. Altogether, we conclude that the SLC34A2-ROS-HIF-1-induced overexpression of EZH2 promotes CRC cells proliferation and chemo-resistance to apoptosis. SLC34A2-ROS-HIF-1-EZH2 signaling pathway might serve as a novel therapeutic target against CRC.

Preclinical activity of lenalidomide in metastatic colorectal cancer.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. e14654-e14654 ◽  
Author(s):  
Valeria Leuci ◽  
Federica Maione ◽  
Maja Todorovic ◽  
Enrico Giraudo ◽  
Loretta Gammaitoni ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Tumor Microenvironment ◽  
Metastatic Colorectal Cancer ◽  
Antitumor Effect ◽  
Average Rate ◽  
Colon Tissue ◽  
Inhibition Of Angiogenesis ◽  
Main Activity

e14654 Background: Metastatic colorectal cancer (mCRC) is currently considered incurable. Recent evidences support the importance of micro-environment and angiogenesis on survival and propagation of mCRC. Lenalidomide is a derivative of thalidomide, effective in several hematologic disorders, whose activity includes modulation on tumor microenvironment and inhibition of angiogenesis. We investigated the preclinical activity of lenalidomide against mCRC in vitro and in vivo. Methods: CRC cell lines and primary mCRC cells were treated with scalar concentrations of lenalidomide (10 to 150 µM). Two cohorts of NOD/SCID mice (n=8) implanted with mCRC xenografts derived from two different patients were treated orally for 21 consecutive days with Lenalidomide (50 mg/Kg/die). Results: In vivo a significant reduction of tumor growth was observed in one cohort compared to untreated controls (n=8), 247±118 mm3 vs. 567±216 mm3 (P=0.02), while no significant differences occurred in the second cohort. Pathology review on tumor samples removed from the mice revealed a trend toward the restoration of normal colon tissue morphology and architecture in lenalidomide treated samples compared to untreated controls. Quantitative evaluation of tumor angiogenesis on the responsive cohort, by immunohistochemistry for factor VIII, revealed a tumor-vessel reduction of 49% with lenalidomide compared to controls. Moreover, necrotic-hypoxic areas were reduced by 6 fold in lenalidomide treated xenografts compared to untreated controls (P<0.001). Treatment in vitro did not result in any direct antitumor effect. The average rate of cell death in treated samples was 15.5±2.1%, 15.3±6.7%, and 11.5±3.4% at lenalidomide doses of 10, 40 and 150 µM respectively, compared to 14.6±2.6% of untreated controls. Conclusions: Our data indicate a potential activity of lenalidomide against mCRC. The beneficial effect was limited in-vivo, supporting the hypothesis that the main activity of lenalidomide is exerted through modulation of tumor microenvironment and angiogenesis rather than a direct antitumor effect. Our findings may be of clinical relevance and support further investigations to explore the benefit of lenalidomide in the challenging setting of mCRC.

scholarly journals In VivoExpression of Streptococcus pyogenes Immunogenic Proteins during Tibial Foreign Body Infection

2014 ◽  
Vol 82 (9) ◽  
pp. 3891-3899 ◽  
Author(s):  
Jeffrey A. Freiberg ◽  
Kevin S. McIver ◽  
Mark E. Shirtliff
Keyword(s):  
Group A Streptococcus ◽  
Rabbit Model ◽  
Growth Conditions ◽  
Rabbit Serum ◽  
Tissue Samples ◽  
Wide Range ◽  
Group A ◽  
Immunogenic Proteins

ABSTRACTGroup A streptococcus (GAS) is an important human pathogen that causes a number of diseases with a wide range of severities. While all known strains of GAS are still sensitive to penicillin, there have been reports of antibiotic treatment failure in as many as 20% to 40% of cases. Biofilm formation has been implicated as a possible cause for these failures. A biofilm is a microbially derived, sessile community where cells grow attached to a surface or as a bacterial conglomerate and surrounded by a complex extracellular matrix. While the ability of group A streptococcus to form biofilms in the laboratory has been shown, there is a lack of understanding of the role of GAS biofilms during an infection. We hypothesized that during infections, GAS exhibits a biofilm phenotype, complete with unique protein expression. To test this hypothesis, a rabbit model of GAS osteomyelitis was developed. A rabbit was inoculated with GAS using an infected indwelling device. Following the infection, blood and tissue samples were collected. Histological samples of the infected tibia were prepared, and the formation of a biofilmin vivowas visualized using peptide nucleic acid fluorescentin situhybridization (PNA-FISH) and confocal microscopy. In addition, Western blotting with convalescent rabbit serum detected cell wall proteins expressedin vitrounder biofilm and planktonic growth conditions. Immunogenic proteins were then identified using matrix-assisted laser desorption ionization–time of flight tandem mass spectrometry (MALDI-TOF/TOF MS). These identities, along with thein vivoresults, support the hypothesis that GAS forms biofilms during an infection. This unique phenotype should be taken into consideration when designing a vaccine or any other treatment for group A streptococcus infections.

scholarly journals The Novel Arylamidine T-2307 Selectively Disrupts Yeast Mitochondrial Function by Inhibiting Respiratory Chain Complexes

2019 ◽  
Vol 63 (8) ◽  
Author(s):  
Kohei Yamashita ◽  
Taiga Miyazaki ◽  
Yoshiko Fukuda ◽  
Junichi Mitsuyama ◽  
Tomomi Saijo ◽  
...  
Keyword(s):  
Respiratory Chain ◽  
Mitochondrial Function ◽  
Yeast Cells ◽  
Dependent Manner ◽  
The Novel ◽  
Respiratory Chain Complexes ◽  
Content Type ◽  
Chain Complexes

ABSTRACT The novel arylamidine T-2307 exhibits broad-spectrum in vitro and in vivo antifungal activities against clinically significant pathogens. Previous studies have shown that T-2307 accumulates in yeast cells via a specific polyamine transporter and disrupts yeast mitochondrial membrane potential. Further, it has little effect on rat liver mitochondrial function. The mechanism by which T-2307 disrupts yeast mitochondrial function is poorly understood, and its elucidation may provide important information for developing novel antifungal agents. This study aimed to determine how T-2307 promotes yeast mitochondrial dysfunction and to investigate the selectivity of this mechanism between fungi and mammals. T-2307 inhibited the respiration of yeast whole cells and isolated yeast mitochondria in a dose-dependent manner. The similarity of the effects of T-2307 and respiratory chain inhibitors on mitochondrial respiration prompted us to investigate the effect of T-2307 on mitochondrial respiratory chain complexes. T-2307 particularly inhibited respiratory chain complexes III and IV not only in Saccharomyces cerevisiae but also in Candida albicans, indicating that T-2307 acts against pathogenic fungi in a manner similar to that of yeast. Conversely, T-2307 showed little effect on bovine respiratory chain complexes. Additionally, we demonstrated that the inhibition of respiratory chain complexes by T-2307 resulted in a decrease in the intracellular ATP levels in yeast cells. These results indicate that inhibition of respiratory chain complexes III and IV is a key factor for selective disruption of yeast mitochondrial function and antifungal activity.

scholarly journals Preclinical In Vitro and In Vivo Evaluation of [18F]FE@SUPPY for Cancer PET Imaging: Limitations of a Xenograft Model for Colorectal Cancer

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
T. Balber ◽  
J. Singer ◽  
N. Berroterán-Infante ◽  
M. Dumanic ◽  
L. Fetty ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Pet Imaging ◽  
Xenograft Model ◽  
Preclinical Study ◽  
Colon Tissue ◽  
In Vivo Evaluation ◽  
Pet Tracer ◽  
Ht 29

Molecular imaging probes such as PET-tracers have the potential to improve the accuracy of tumor characterization by directly visualizing the biochemical situation. Thus, molecular changes can be detected early before morphological manifestation. The A3 adenosine receptor (A3AR) is described to be highly expressed in colon cancer cell lines and human colorectal cancer (CRC), suggesting this receptor as a tumor marker. The aim of this preclinical study was the evaluation of F18FE@SUPPY as a PET-tracer for CRC using in vitro imaging and in vivo PET imaging. First, affinity and selectivity of FE@SUPPY and its metabolites were determined, proving the favorable binding profile of FE@SUPPY. The human adenocarcinoma cell line HT-29 was characterized regarding its hA3AR expression and was subsequently chosen as tumor graft. Promising results regarding the potential of F18FE@SUPPY as a PET-tracer for CRC imaging were obtained by autoradiography as ≥2.3-fold higher accumulation of F18FE@SUPPY was found in CRC tissue compared to adjacent healthy colon tissue from the same patient. Nevertheless, first in vivo studies using HT-29 xenografts showed insufficient tumor uptake due to (1) poor conservation of target expression in xenografts and (2) unfavorable pharmacokinetics of F18FE@SUPPY in mice. We therefore conclude that HT-29 xenografts are not adequate to visualize hA3ARs using F18FE@SUPPY.

scholarly journals Selection of a Malignant Subpopulation from a Colorectal Cancer Cell Line

2019 ◽  
Author(s):  
Pei-Lun Lai ◽  
Ting-Chun Chen ◽  
Chun-Yen Feng ◽  
Hsuan Lin ◽  
Ng Wu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Line ◽  
Prognostic Biomarker ◽  
Differentially Expressed Gene ◽  
Cancer Cell Line ◽  
Experimental Models ◽  
Colorectal Cancer Cell Line ◽  
Selection Of

AbstractColorectal cancer (CRC) is a leading cause of death from cancer worldwide. Thus, there is an emerging need for new experimental models that allow identification and validation of biomarkers for CRC-specific progression. In this study, we propose a repeated sphere-forming assay as a strategy to select a malignant subpopulation from a CRC line, HCT116. We validated our assay by confirming that three canonical stemness markers, Nanog, Oct4, and Lgr5, were up-regulated in the sphere state at every generation of the selection assay. The resulting line, after eight rounds of selection, exhibited an increased sphere-forming capacityin vitroand tumorgenicityin vivo. Furthermore, dipeptidase 1 (DPEP1) was identified as the major differentially expressed gene in the selected clone, and depletion of DPEP1 suppressed the elevated sphere-forming capacityin vitroand tumorgenicityin vivo. Overall, we have established an experimental strategy for the isolation of a malignant subpopulation from a CRC cell line. Results from our model also suggested that DPEP1 can serve as a promising prognostic biomarker for CRC.

scholarly journals Antiproliferative Effect of Aspirin on Colorectal Cancer Cell Line

2018 ◽  
Vol 12 (5) ◽  
pp. 1-4
Author(s):  
Malihe Bagheri ◽  
◽  
Amir Reza Hesari ◽  
Parisa Zia Sarabi ◽  
Hamid Reza Rahimi ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell Line ◽  
Sw480 Cell ◽  
Colorectal Cancer Cell Line ◽  
Range Analysis ◽  
Cytotoxic Effects

Background: Nonsteroidal anti-inflammatory drugs (NSAIDs) such as Aspirin may have anticancer properties, and can be effective as a novel strategy for the treatment of colorectal cancer (CRC). The aim of this study was to assess the cytotoxic effects of Aspirin drug in CRC cell lines compared with Oxaliplatin drug in vitro. Methods: Cell viability was assessed after treatment of SW742 and SW480 cells with Aspirin and Oxaliplatin by MTT assay, and the amount of IC50 was determined. Statistical analysis was performed through one-way ANOVA and Tukey multiple range analysis (SPSS 19.0 software (P <0.05). Results: Aspirin and Oxaliplatin considerably inhibited the growth of SW742 and SW480 cell lines. SW742 cell line was more sensitive to Aspirin than SW480 cell line. The cytotoxic effect of Oxaliplatin was higher than Aspirin in both cell lines. Conclusions: This study demonstrated that both Aspirin and Oxaliplatin have cytotoxic effects on SW742 and SW480 cell lines in vitro. Thus, Aspirin may be considered as a therapeutic agent in CRC, however, further in vivo investigations are required to fully establish this effect.

Sign in / Sign up

Export Citation Format

Share Document