scholarly journals Gels for Live Analysis of Compartmentalized Environments (GLAnCE): A Tissue Model to Probe Tumour Phenotypes at Tumour-Stroma Interfaces

2019 ◽  
Author(s):  
Elisa D’Arcangelo ◽  
Nila C. Wu ◽  
Tianhao Chen ◽  
Andi Shahaj ◽  
Jose L. Cadavid ◽  
...  
Keyword(s):  
Tumour Cell ◽  
Cell Movement ◽  
Cell Interaction ◽  
Carcinoma Cells ◽  
Tumour Stroma ◽  
Paracrine Signaling ◽  
Tumour Cell Invasion ◽  
A Site ◽  
Spatial Stratification

AbstractThe interface between a tumour and the adjacent stroma is a site of great importance for tumour development. At this site, carcinoma cells are highly proliferative, undergo invasive phenotypic changes, and directly interact with surrounding stromal cells, such as cancer-associated fibroblasts (CAFs) which further exert pro-tumorigenic effects. Here we describe the development of GLAnCE (Gels for Live Analysis of Compartmentalized Environments), an easy-to-use hydrogel-culture platform for investigating CAF-tumour cell interaction dynamics in vitro at a tumour-stroma interface. GLAnCE enables observation of CAF-mediated enhancement of both tumour cell proliferation and invasion at the tumour-stroma interface in real time, as well as stratification between phenotypes at the interface versus in the bulk tumour tissue compartment. We found that CAF presence resulted in the establishment of an invasion-permissive, interface-specific matrix environment, that leads to carcinoma cell movement outwards from the tumour edge and tumour cell invasion. Furthermore, the spatial stratification capability of GLAnCE was leveraged to discern differences between tumour cell epithelial-to-mesenchymal (EMT) transition genes induced by paracrine signaling from CAFs versus genes induced by interface-specific, CAF-mediated microenvironment. GLAnCE combines high usability and tissue complexity, to provide a powerful in vitro platform to probe mechanisms of tumour cell movement specific to the microenvironment at the tumour-stroma interface.

scholarly journals CTEN Induces Tumour Cell Invasion and Survival and Is Prognostic in Radiotherapy-Treated Head and Neck Cancer

Cancers ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 2963
Author(s):  
Jason C. Fleming ◽  
Jeongmin Woo ◽  
Karwan Moutasim ◽  
Christopher J. Hanley ◽  
Steven J. Frampton ◽  
...  
Keyword(s):  
Head And Neck ◽  
Cell Invasion ◽  
Tumour Cell ◽  
3D Culture ◽  
Response To Therapy ◽  
Antibody Array ◽  
Hpv Status ◽  
Tumour Cell Invasion

Head and neck squamous cell carcinoma (HNSCC) is a heterogenous disease treated with surgery and/or (chemo) radiotherapy, but up to 50% of patients with late-stage disease develop locoregional recurrence. Determining the mechanisms underpinning treatment resistance could identify new therapeutic targets and aid treatment selection. C-terminal tensin-like (CTEN) is a member of the tensin family, upregulated in several cancers, although its expression and function in HNSCC are unknown. We found that CTEN is commonly upregulated in HNSCC, particularly HPV−ve tumours. In vitro CTEN was upregulated in HPV−ve (n = 5) and HPV+ve (n = 2) HNSCC cell lines. Stable shRNA knockdown of CTEN in vivo significantly reduced tumour growth (SCC-25), and functional analyses in vitro showed that CTEN promoted tumour cell invasion, colony formation and growth in 3D-culture (SCC-25, Detroit 562). RNA sequencing of SCC-25 cells following CTEN siRNA knockdown identified 349 differentially expressed genes (logFC > 1, p < 0.05). Gene ontology analysis highlighted terms relating to cell locomotion and apoptosis, consistent with in vitro findings. A membrane-based antibody array confirmed that CTEN regulated multiple apoptosis-associated proteins, including HSP60 and cleaved caspase-3. Notably, in a mixed cohort of HPV+ve and HPV−ve HNSCC patients (n = 259), we found a significant, independent negative association of CTEN with prognosis, limited to those patients treated with (chemo)radiotherapy, not surgery, irrespective of human papillomavirus (HPV) status. These data show that CTEN is commonly upregulated in HNSCC and exerts several functional effects. Its potential role in modulating apoptotic response to therapy suggests utility as a predictive biomarker or radio-sensitising target.

A New In-Vitro Model to Study Tumour Cell Invasion

Metastasis ◽  
1980 ◽  
pp. 28-32 ◽  
Author(s):  
R. Tchao ◽  
A. B. Schleich ◽  
M. Frick ◽  
A. Mayer
Keyword(s):  
Cell Invasion ◽  
Tumour Cell ◽  
In Vitro Model ◽  
Tumour Cell Invasion ◽  
Vitro Model

TUMOUR CELL MOVEMENT DURING HEATING AND HUMIDIFICATION OF INSUFFLATING CO2: AN IN VITRO MODEL

1998 ◽  
Vol 68 (10) ◽  
pp. 740-742 ◽  
Author(s):  
Michael L. Texler ◽  
Grant King ◽  
Peter J. Hewett
Keyword(s):  
Tumour Cell ◽  
In Vitro Model ◽  
Cell Movement ◽  
Vitro Model

CBIO-17. A NOVEL ROLE FOR ATR KINASE DETERMINES GLIOBLASTOMA TUMOUR CELL INVASION

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi30-vi30
Author(s):  
ross carruthers ◽  
Sarah Derby ◽  
Karen Strathdee ◽  
Anthony Chalmers ◽  
Jim Norman ◽  
...  
Keyword(s):  
Dna Damage ◽  
Cell Invasion ◽  
Tumour Cell ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Tumour Cell Invasion ◽  
Atr Kinase

Abstract BACKGROUND: Widespread contamination of the brain with malignant cells is a predominant feature of glioblastoma (GBM) and fatal brainstem infiltration is frequently observed at autopsy. Whilst radiotherapy improves survival, irradiation increases GBM cell invasion, resulting in sublethal dose to cells migrating outside the irradiated volume. Tumour cell invasion should be a therapeutic priority if survival is to be improved. The responsible molecular mechanisms are key to improving outcomes but remain enigmatic. Ataxia telangiectasia and rad3-related (ATR) is a DNA damage response (DDR) kinase involved in DNA replication stress (RS) response and is an established therapeutic target for GBM. In this study we demonstrate a novel role for ATR kinase in facilitating malignant cell invasion. METHODS AND RESULTS: Invading margins of human GBM samples demonstrated increased pATR expression relative to core. Live cell imaging demonstrated a reduction in cell velocity following ATR inhibition (ATRi; VE822) or ATR siRNA, and a retraction defect was evident in vitro. Extensive cytoplasmic vacuolation occurred following ATRi or siRNA which were single walled structures on electron microscopy which could engulf high molecular weight dextran, suggesting blockade of macropinosome processing. Live cell imaging with GFP-integrin α5 and integrin recycling assays showed integrin sequestration within macropinosomes and reduced integrin internalisation respectively. Interrogation of a published ‘ATR interactome’ revealed ATR targets with functions in endocytic vesicle trafficking. Intravital in vivo imaging of murine xenograft tumours confirmed vacuolation and dextran uptake following ATRi, whilst a further study demonstrated reduced invading tumour cells following ATRi in intracranial xenografts. CONCLUSION: We demonstrate a novel role for ATR in facilitating macropinocytic vesicle trafficking and integrin internalisation. ATRi results in a profound motility defect in vitro and in vivo. ATR inhibitors are entering early phase trials as radiation sensitisers and we propose that therapeutic benefit will extend beyond DNA damage potentiation.

Significant Down-Regulation of BAALC during Lineage Specific Hematopoietic Differentiation.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4189-4189
Author(s):  
Claudia D. Baldus ◽  
Martina Komor ◽  
Dieter Hoelzer ◽  
Albert de la Chapelle ◽  
Eckhard Thiel ◽  
...  
Keyword(s):  
Expression Profiling ◽  
Cell Movement ◽  
Cell Interaction ◽  
Hematopoietic Differentiation ◽  
Rt Pcr ◽  
Down Regulation ◽  
Gm Csf ◽  
Specific Differentiation ◽  
Cd34 Cells

Abstract The human gene BAALC (Brain And Acute Leukemia, Cytoplasmic) implicated in normal hematopoiesis and leukemia is a marker of immature hematopoietic progenitor cells (PNAS2001;98:13901). To explore its role in hematopoiesis we investigated the regulation of BAALC during lineage specific differentiation. Semi-quantitative RT-PCR was performed on subsets of in vitro differentiated bone marrow CD34+ cells from healthy individuals using the cytokines G-CSF, M-CSF or EPO. Cultures were harvested on day 4, 8, 12, and 16. In addition, oligonucleotide microarray analyses (HG-U133A, Affymetrix) of in vitro stimulated CD34+ cells were independently carried out using EPO, TPO, G/GM-CSF to induce lineage-specific differentiation. For each of the lineages, cells were harvested at days 4, 7 and 11 for expression profiling. All experiments were done in triplicates for each time point and condition. RT-PCR revealed down-regulation of BAALC and CD34 as early as day 4 in cultures with G-CSF, M-CSF or EPO. In concordance, gene expression profiling showed significant down-regulation of BAALC and CD34 on day 4 of culture (BAALC: EPO p=0.04, G/GM-CSF p=0.17, TPO p=0.03; CD34: EPO p=0.01, G/GM-CSF p=0.003, TPO p=0.01). 275 genes showing a significant correlation (correlation coefficient r&gt;0.95) to BAALC expression levels (at the 4 time points and using 3 different cytokines) were identified. Of these CD34 ranked second (r=0.99), and genes related to leukemia and neuronal cells included: Hepatic leukemia factor (HLF; r=0.99) and Paired box gene 5 (PAX5; r=0.99). It has been speculated that BAALC might be involved in cell movement or cell-cell interaction; we therefore investigated the association to genes implicated in cytoskeletal function. Expression of Myosin 5C (MYO5C; r=0.99) and Synaptic nulcei expressed gene 2 (SYNE2; r=0.95) were highly correlated with BAALC expression across all conditions (EPO, TPO, G/GM-CSF). In conclusion, using two independent approaches we show the significant down-regulation of BAALC during hematopoietic differentiation. This underscores the potential role of BAALC in early hematopoiesis and the possible contribution of aberrant BAALC expression in leukemogenesis. We postulate that similar to CD34, PAX, and HLF implicated in hematopoiesis and leukemia, down-regulation of BAALC and structural genes including Myosin 5C are critical steps during hematopoietic differentiation.

scholarly journals Fibronectin enhances in vitro monocyte-macrophage-mediated tumoricidal activity

Blood ◽  
1982 ◽  
Vol 60 (2) ◽  
pp. 430-435 ◽  
Author(s):  
RT Perri ◽  
NE Kay ◽  
J McCarthy ◽  
RL Vessella ◽  
HS Jacob ◽  
...  
Keyword(s):  
Human Tumor ◽  
Human Monocyte ◽  
Cell Interaction ◽  
Carcinoma Cells ◽  
Target Cells ◽  
Tumoricidal Activity ◽  
Dose Dependent Fashion ◽  
Human Fibronectin ◽  
Dose Dependent

Abstract Control of neoplastic proliferation reflects in part monocyte/macrophage destruction of target cells--destruction that evidently requires cell-cell interaction. We herein show it to involve the natural plasma opsonin, fibronectin. With two cultured human tumor lines--Malme melanoma and CAK-I renal carcinoma cells--addition of fibronectin, purified to homogeneity, enhances macrophage-mediated cytotoxicity 2--4-fold (p less than 0.01). Both fresh human monocytes or the U-937-cultured macrophage line become more lethal to tumor cells with added fibronectin. The fibronectin-enhanced monocyte and U-937 tumoricidal activity occurred in a dose-dependent fashion. Specificity of fibronectin's action was validated by use of affinity-purified rabbit antifibronectin antibody, which completely abated its enhancement of tumoricidal activity. Enhancement of tumoricidal activity did not occur when monocytes or U-937 were exposed to fibronectin-coated plates. However, the addition of soluble fibronectin to fibronectin-coated plates was then capable of enhancing cytocidal activity. These studies demonstrate that human fibronectin is capable of increasing both fresh and cultured human monocyte tumor-directed cytotoxicity. Fibronectin appears to be a potentially important circulating molecule that may favorably influence human monocyte tumor cell cytotoxicity.

scholarly journals Discovery of a small-molecule inhibitor of specific serine residue BAD phosphorylation

2018 ◽  
Vol 115 (44) ◽  
pp. E10505-E10514 ◽  
Author(s):  
Vijay Pandey ◽  
Baocheng Wang ◽  
Chakrabhavi Dhananjaya Mohan ◽  
Ainiah Rushdiana Raquib ◽  
Shobith Rangappa ◽  
...  
Keyword(s):  
Small Molecule ◽  
Human Cancer ◽  
Regulatory Protein ◽  
Single Agent ◽  
Xenograft Model ◽  
Specific Inhibitor ◽  
Carcinoma Cells ◽  
Site Specific ◽  
A Site

Human BCL-2–associated death promoter (hBAD) is an apoptosis-regulatory protein mediating survival signals to carcinoma cells upon phosphorylation of Ser99, among other residues. Herein, we screened multiple small-molecule databases queried in a Laplacian-modified naive Bayesian-based cheminformatics platform and identified a Petasis reaction product as a site-specific inhibitor for hBAD phosphorylation. Based on apoptotic efficacy against mammary carcinoma cells, N-cyclopentyl-3-((4-(2,3-dichlorophenyl) piperazin-1-yl) (2-hydroxyphenyl) methyl) benzamide (NPB) was identified as a potential lead compound. In vitro biochemical analyses demonstrated that NPB inhibited the phosphorylation of hBAD specifically on Ser99. NPB was observed to exert this effect independently of AKT and other kinase activities despite the demonstration of AKT-mediated BAD-Ser99 phosphorylation. Using a structure-based bioinformatics platform, we observed that NPB exhibited predicted interactions with hBAD in silico and verified the same by direct binding kinetics. NPB reduced phosphorylation of BAD-Ser99 and enhanced caspase 3/7 activity with associated loss of cell viability in various human cancer cell lines derived from mammary, endometrial, ovarian, hepatocellular, colon, prostatic, and pancreatic carcinoma. Furthermore, by use of a xenograft model, it was observed that NPB, as a single agent, markedly diminished BAD phosphorylation in tumor tissue and significantly inhibited tumor growth. Similar doses of NPB utilized in acute toxicity studies in mice did not exhibit significant effects. Hence, we report a site-specific inhibitor of BAD phosphorylation with efficacy in tumor models.

scholarly journals Fibronectin enhances in vitro monocyte-macrophage-mediated tumoricidal activity

Blood ◽  
1982 ◽  
Vol 60 (2) ◽  
pp. 430-435 ◽  
Author(s):  
RT Perri ◽  
NE Kay ◽  
J McCarthy ◽  
RL Vessella ◽  
HS Jacob ◽  
...  
Keyword(s):  
Human Tumor ◽  
Human Monocyte ◽  
Cell Interaction ◽  
Carcinoma Cells ◽  
Target Cells ◽  
Tumoricidal Activity ◽  
Dose Dependent Fashion ◽  
Human Fibronectin ◽  
Dose Dependent

Control of neoplastic proliferation reflects in part monocyte/macrophage destruction of target cells--destruction that evidently requires cell-cell interaction. We herein show it to involve the natural plasma opsonin, fibronectin. With two cultured human tumor lines--Malme melanoma and CAK-I renal carcinoma cells--addition of fibronectin, purified to homogeneity, enhances macrophage-mediated cytotoxicity 2--4-fold (p less than 0.01). Both fresh human monocytes or the U-937-cultured macrophage line become more lethal to tumor cells with added fibronectin. The fibronectin-enhanced monocyte and U-937 tumoricidal activity occurred in a dose-dependent fashion. Specificity of fibronectin's action was validated by use of affinity-purified rabbit antifibronectin antibody, which completely abated its enhancement of tumoricidal activity. Enhancement of tumoricidal activity did not occur when monocytes or U-937 were exposed to fibronectin-coated plates. However, the addition of soluble fibronectin to fibronectin-coated plates was then capable of enhancing cytocidal activity. These studies demonstrate that human fibronectin is capable of increasing both fresh and cultured human monocyte tumor-directed cytotoxicity. Fibronectin appears to be a potentially important circulating molecule that may favorably influence human monocyte tumor cell cytotoxicity.

In vitro FTIR microspectroscopy analysis of primary oral squamous carcinoma cells treated with cisplatin and 5-fluorouracil: a new spectroscopic approach for studying the drug–cell interaction

The Analyst ◽  
2018 ◽  
Vol 143 (14) ◽  
pp. 3317-3326 ◽  
Author(s):  
Elisabetta Giorgini ◽  
Simona Sabbatini ◽  
Romina Rocchetti ◽  
Valentina Notarstefano ◽  
Corrado Rubini ◽  
...  
Keyword(s):  
Squamous Carcinoma ◽  
Cell Interaction ◽  
Carcinoma Cells ◽  
Ftir Microspectroscopy ◽  
Squamous Carcinoma Cells ◽  
Oral Squamous Carcinoma

In vitro FTIRM analysis of primary OSCCs treated with cisplatin and 5-fluorouracil for the drug–cell interaction.

Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

1974 ◽  
Vol 32 ◽  
pp. 320-321
Author(s):  
J. P. Revel
Keyword(s):  
Developmental Stages ◽  
Three Dimensional ◽  
Cell Movement ◽  
Cellular Interactions ◽  
Serial Sections ◽  
Cell Sheets ◽  
Freeze Etching ◽  
Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

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