scholarly journals Cellular Integrin α5β1 and Exosomal ADAM17 Mediate the Binding and Uptake of Exosomes Produced by Colorectal Carcinoma Cells

2021 ◽  
Vol 22 (18) ◽  
pp. 9938
Author(s):  
Beatriz Cardeñes ◽  
Irene Clares ◽  
Víctor Toribio ◽  
Lucía Pascual ◽  
Soraya López-Martín ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Cancer Progression ◽  
Peritoneal Metastasis ◽  
Peritoneal Dissemination ◽  
Mesothelial Cell ◽  
Surface Proteins ◽  
Size Exclusion ◽  
Integrin Α5β1 ◽  
Peritoneal Mesothelial Cells ◽  
Exclusion Chromatography

Approximately 25% of colorectal cancer (CRC) patients develop peritoneal metastasis, a condition associated with a bleak prognosis. The CRC peritoneal dissemination cascade involves the shedding of cancer cells from the primary tumor, their transport through the peritoneal cavity, their adhesion to the peritoneal mesothelial cells (PMCs) that line all peritoneal organs, and invasion of cancer cells through this mesothelial cell barrier and underlying stroma to establish new metastatic foci. Exosomes produced by cancer cells have been shown to influence many processes related to cancer progression and metastasis. In epithelial ovarian cancer these extracellular vesicles (EVs) have been shown to favor different steps of the peritoneal dissemination cascade by changing the functional phenotype of cancer cells and PMCs. Little is currently known, however, about the roles played by exosomes in the pathogenesis and peritoneal metastasis cascade of CRC and especially about the molecules that mediate their interaction and uptake by target PMCs and tumor cells. We isolated exosomes by size−exclusion chromatography from CRC cells and performed cell-adhesion assays to immobilized exosomes in the presence of blocking antibodies against surface proteins and measured the uptake of fluorescently-labelled exosomes. We report here that the interaction between integrin α5β1 on CRC cells (and PMCs) and its ligand ADAM17 on exosomes mediated the binding and uptake of CRC-derived exosomes. Furthermore, this process was negatively regulated by the expression of tetraspanin CD9 on exosomes.

EWI-2 modulates lymphocyte integrin α4β1 functions

Blood ◽  
2004 ◽  
Vol 103 (8) ◽  
pp. 3013-3019 ◽  
Author(s):  
Tatiana V. Kolesnikova ◽  
Christopher S. Stipp ◽  
Ravi M. Rao ◽  
William S. Lane ◽  
Francis W. Luscinskas ◽  
...  
Keyword(s):  
Cell Surface ◽  
Cell Adhesion Molecule ◽  
Apparent Size ◽  
Surface Proteins ◽  
Size Exclusion ◽  
Leukemia Cells ◽  
Wild Type ◽  
Vascular Cell Adhesion ◽  
Exclusion Chromatography ◽  
Cell Adhesion Molecule 1

Abstract The most prominent cell-surface integrin α4β1 partner, a 70-kDa protein, was isolated from MOLT-4 T leukemia cells, using anti–α4β1 integrin antibody-coated beads. By mass spectrometry, this protein was identified as EWI-2, a previously described cell-surface partner for tetraspanin proteins CD9 and CD81. Wild-type EWI-2 overexpression had no effect on MOLT-4 cell tethering and adhesion strengthening on the α4β1 ligand, vascular cell adhesion molecule-1 (VCAM-1), in shear flow assays. However, EWI-2 markedly impaired spreading and ruffling on VCAM-1. In contrast, a mutant EWI-2 molecule, with a different cytoplasmic tail, neither impaired cell spreading nor associated with α4β1 and CD81. The endogenous wild-type EWI-2–CD81–α4β1 complex was fully soluble, and highly specific as seen by the absence of other MOLT-4 cell-surface proteins. Also, it was relatively small in size (0.5 × 106 Da to 4 × 106 Da), as estimated by size exclusion chromatography. Overexpression of EWI-2 in MOLT-4 cells caused reorganization of cell-surface CD81, increased the extent of CD81-CD81, CD81-α4β1, and α4β1-α4β1 associations, and increased the apparent size of CD81-α4β1 complexes. We suggest that EWI-2–dependent reorganization of α4β1-CD81 complexes on the cell surface is responsible for EWI-2 effects on integrin-dependent morphology and motility functions. (Blood. 2004;103: 3013-3019)

scholarly journals Production of IL1-beta by ovarian cancer cells induces mesothelial cell beta1-integrin expression facilitating peritoneal dissemination

2012 ◽  
Vol 5 (1) ◽  
pp. 7 ◽  
Author(s):  
Takafumi Watanabe ◽  
Toshihiro Hashimoto ◽  
Takashi Sugino ◽  
Shu Soeda ◽  
Hiroshi Nishiyama ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cancer Cells ◽  
Peritoneal Dissemination ◽  
Mesothelial Cell ◽  
Ovarian Cancer Cells ◽  
Integrin Expression ◽  
Beta1 Integrin

scholarly journals Gastric cancer derived exosomes induce peritoneal mesothelial cell EMT through TGF-β1/Smads pathway to promote peritoneal metastasis

2021 ◽  
Author(s):  
Jungang Dong ◽  
Zhongbo Zhu ◽  
Guoning Cui ◽  
Zhixuan Zhang ◽  
Juan Yue ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Peritoneal Metastasis ◽  
Epithelial Mesenchymal Transition ◽  
Critical Role ◽  
Mesothelial Cell ◽  
Mesothelial Cells ◽  
Mesenchymal Transition ◽  
Peritoneal Mesothelial Cells ◽  
Metastatic Microenvironment ◽  
Tgf Β1

Epithelial-mesenchymal transition (EMT) plays an important role in peritoneal metastasis of Gastric cancer (GC). Tumor exosomes can mediate tumor directed metastasis, and TGF-β1 is an important factor in inducing tumor Epithelial mesenchymal transition. However, it is not clear whether GC derived exosomes can induce peritoneal mesothelial cells through the TGF-β1/ Smads pathway and the effect of injured peritoneal mesothelial cells on the biological characteristics of GC cells. In this study, we demonstrated that GC-derived exosomes can activate the TGF-β1/Smads pathway in peritoneal mesothelial cells and induce the corresponding EMT process, and that the injured peritoneal mesothelial cells can improve the migration and adhesion of GC cells. Taken together, these data further support the critical role of exosomes in the remodeling of the pre-metastatic microenvironment.

scholarly journals Size-Exclusion Chromatography as a Technique for the Investigation of Novel Extracellular Vesicles in Cancer

Cancers ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3156
Author(s):  
Daniel S. K. Liu ◽  
Flora M. Upton ◽  
Eleanor Rees ◽  
Christopher Limb ◽  
Long R. Jiao ◽  
...  
Keyword(s):  
Systematic Review ◽  
Cancer Cells ◽  
Size Exclusion Chromatography ◽  
Extracellular Vesicles ◽  
Biomarker Discovery ◽  
Extracellular Vesicle ◽  
Size Exclusion ◽  
Isolation Method ◽  
Isolation And Purification ◽  
Exclusion Chromatography

Cancer cells release extracellular vesicles, which are a rich target for biomarker discovery and provide a promising mechanism for liquid biopsy. Size-exclusion chromatography (SEC) is an increasingly popular technique, which has been rediscovered for the purposes of extracellular vesicle (EV) isolation and purification from diverse biofluids. A systematic review was undertaken to identify all papers that described size exclusion as their primary EV isolation method in cancer research. In all, 37 papers were identified and discussed, which showcases the breadth of applications in which EVs can be utilised, from proteomics, to RNA, and through to functionality. A range of different methods are highlighted, with Sepharose-based techniques predominating. EVs isolated using SEC are able to identify cancer cells, highlight active pathways in tumourigenesis, clinically distinguish cohorts, and remain functionally active for further experiments.

scholarly journals Use of photoimmunoconjugates to characterize ABCB1 in cancer cells

Nanophotonics ◽  
2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Barry J. Liang ◽  
Sabrina Lusvarghi ◽  
Suresh V. Ambudkar ◽  
Huang-Chiao Huang
Keyword(s):  
Cancer Cells ◽  
Triple Negative ◽  
Size Exclusion ◽  
Detection Methods ◽  
Drug Resistant ◽  
Drug Transporter ◽  
Open Conformation ◽  
Exclusion Chromatography ◽  
Insight Into

Abstract Accurate detection of ATP-binding cassette drug transporter ABCB1 expression is imperative for precise identification of drug-resistant tumors. Existing detection methods fail to provide the necessary molecular details regarding the functional state of the transporter. Photoimmunoconjugates are a unique class of antibody–dye conjugates for molecular diagnosis and therapeutic treatment. However, conjugating hydrophobic photosensitizers to hydrophilic antibodies is quite challenging. Here, we devise a photoimmunoconjugate that combines a clinically approved benzoporphyrin derivative (BPD) photosensitizer and the conformational-sensitive UIC2 monoclonal antibody to target functionally active human ABCB1 (i.e., ABCB1 in the inward-open conformation). We show that PEGylation of UIC2 enhances the BPD conjugation efficiency and reduces the amount of non-covalently conjugated BPD molecules by 17%. Size exclusion chromatography effectively separates the different molecular weight species found in the UIC2–BPD sample. The binding of UIC2–BPD to ABCB1 was demonstrated in lipidic nanodiscs and ABCB1-overexpressing triple negative breast cancer (TNBC) cells. UIC2–BPD was found to retain the conformation sensitivity of UIC2, as the addition of ABCB1 modulators increases the antibody reactivity in vitro. Thus, the inherent fluorescence capability of BPD can be used to label ABCB1-overexpressing TNBC cells using UIC2–BPD. Our findings provide insight into conjugation of hydrophobic photosensitizers to conformation-sensitive antibodies to target proteins expressed on the surface of cancer cells.

scholarly journals Macrophage-Derived Small Exosomes: Efficient Transmission and Cytotoxicity to Cancer Cells

2021 ◽  
Author(s):  
Yanyin Lu ◽  
Takanori Eguchi ◽  
Chiharu Sogawa ◽  
Eman A. Taha ◽  
Manh Tien Tran ◽  
...  
Keyword(s):  
Oral Cancer ◽  
Cancer Cells ◽  
Culture System ◽  
Size Exclusion ◽  
Fluorescence Labeling ◽  
Monocytic Cells ◽  
Beta Actin ◽  
Exclusion Chromatography ◽  
Oral Cancer Cells ◽  
Efficient Transmission

Tumor-associated macrophages are a key component in the tumor microenvironment, secreting extracellular vesicles (EVs) such as exosomes and other various factors for intercellular communication. However, macrophage-derived EVs heterogeneity and their cytotoxicity to cancer cells has not been well understood. Here, we aimed to separately isolate various types of macro-phage-EVs by size exclusion chromatography (SEC) method and investigate EV transmission and cytotoxicity to oral cancer cells. For fluorescence-labeling of cellular and EV membranes, palmitoylation signal-fused GFP and tdTomato were expressed in THP-1 monocytic cells and HSC-3 oral cancer cells, respectively. We found that fluorescence-labeled EVs secreted by macrophages were highly transmissive to oral cancer cells than those from parental monocytic cells. In a co-culture system and conditioned medium (CM), a macrophage-secreted unidentified factor was cytotoxic to oral cancer cells. We fractionated macrophage-derived EVs by the SEC method and performed western blotting to characterize various EV types. Three fractions were characterized: small exosomes (EXO-S: < 50 nm) fraction containing HSP90α, HSP90β, CD63 (EV marker) and β-actin; large exosomes (EXO-L: 50-200 nm) fraction containing CD9 (EV marker) and HSP90β; large EVs (100-500 nm) fraction. Notably, the macrophage-derived small exosomes fraction was cytotoxic to oral cancer cells, while large exosomes and large EVs were not. There-fore, it was implicated that macrophage-derived small exosomes are cytotoxic with high trans-mission potential to cancer cells.

scholarly journals Platelets and extracellular vesicles and their cross-talk with cancer

Blood ◽  
2021 ◽  
Author(s):  
Sophia Lazar ◽  
Lawrence E. Goldfinger
Keyword(s):  
Cancer Cells ◽  
Extracellular Vesicles ◽  
Cancer Progression ◽  
Second Messengers ◽  
Inflammatory Cells ◽  
Surface Proteins ◽  
Non Coding Rnas ◽  
Promote Disease Progression ◽  
Mature Mrnas ◽  
Heterogeneous Size

Platelets play significant and varied roles in cancer progression, as detailed throughout this review series, via direct interactions with cancer cells as well as by long-range indirect interactions mediated by platelet releasates. Microvesicles (MV, also referred to as microparticles) released from activated platelets have emerged as major contributors to the platelet-cancer nexus. Interactions of platelet-derived MV (PMV) with cancer cells can promote disease progression through multiple mechanisms, but PMV also harbor anti-tumor functions. This complex relationship derives from the abilities of PMV both to bind to cancer cells as well as to non-transformed cells in the tumor microenvironment, and to transfer platelet-derived contents to the target cell, each of which can have stimulatory or modulatory effects. MV are extracellular vesicles of heterogeneous size, ranging from 100 nm to 1 µm in diameter, shed by living cells by outward budding of the plasma membrane, entrapping local cytosolic contents in an apparently stochastic manner. Hence, PMV are encapsulated by a lipid bilayer harboring surface proteins and lipids mirroring the platelet exterior, with internal components including platelet-derived mature mRNAs and pre-mRNAs, microRNAs (miRNAs) and other non-coding RNAs, proteins, second messengers, and mitochondria. Each of these elements engages in established and putative PMV functions in cancer. In addition, PMV contribute to cancer co-morbidities due to their roles in coagulation and thrombosis, and via interactions with inflammatory cells. However, separating effects of PMV from those of platelets in cancer contexts continues to be a major hurdle (Figure 1). This review will summarize our emerging understanding of the complex roles of PMV in the development and progression of cancer and cancer co-morbidities.

scholarly journals Malignant Ascites Promote Adhesion of Ovarian Cancer Cells to Peritoneal Mesothelium and Fibroblasts

2021 ◽  
Vol 22 (8) ◽  
pp. 4222
Author(s):  
Paweł Uruski ◽  
Justyna Mikuła-Pietrasik ◽  
Martyna Pakuła ◽  
Sylwia Budkiewicz ◽  
Marcin Drzewiecki ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cancer Cells ◽  
Cancer Progression ◽  
Cancer Metastasis ◽  
Malignant Ascites ◽  
Ovarian Cancer Cells ◽  
Vimentin Expression ◽  
Peritoneal Mesothelial Cells ◽  
Peritoneal Mesothelium ◽  
Tgf Β1

Although malignant ascites (MAs) are known to contribute to various aspects of ovarian cancer progression, knowledge regarding their role in the adhesion of cancer cells to normal peritoneal cells is incomplete. Here, we compared the effect of MAs and benign ascites (BAs) on the adhesion of A2780 and OVCAR-3 cancer cells to omentum-derived peritoneal mesothelial cells (PMCs) and peritoneal fibroblasts (PFBs). The results showed that MAs stimulated the adhesion of A2780 and OVCAR-3 cells to PMCs and PFBs more efficiently than did BAs, and the strongest binding occurred when both cancer and normal cells were exposed to the fluid. Intervention studies showed that MAs-driven adhesion of A2780 cells to PMCs/PFBs depends on the presence of TGF-β1 and HGF, whereas binding of OVCAR-3 cells was mediated by TGF-β1, GRO-1, and IGF-1. Moreover, MAs upregulated α5β1 integrin expression on PFBs but not on PMCs or cancer cells, vimentin expression in all cells tested, and ICAM-1 only in cancer cells. When integrin-linked kinase was neutralized in PMCs or PFBs, cancer cell adhesion to PMCs and PFBs decreased. Collectively, our report shows that MAs may contribute to the early stages of ovarian cancer metastasis by modulating the proadhesive interplay between normal and cancer cells.

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