Extracellular Vesicles Play an Important Role in Intercellular Communication Between Bone Marrow Stroma and Hematopoietic Progenitor Cells in Myeloproliferative Neoplasms
Teresa Lopes RamosLuis Ignacio Sánchez‐AbarcaBeatriz RosónAlba RedondoConcepción RodríguezHaidy RodríguezRebeca OrtegaAna Isabel RicoSilvia PreciadoOlga López VillarSandra MuntiónJosé Ramón González‐PorrasJesús María Hernández‐RivasJavier De Las RivasÁngel Hernández‐HernándezFermín Sánchez‐GuijoConsuelo del Cañizo
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CD90
Extracellular vesicles
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Abstract Exosomes and microvesicles are two classes of submicroscopic vesicle released by cells into the extracellular space. Collectively referred to as extracellular vesicles, these membrane containers facilitate important cell–cell communication by carrying a diverse array of signaling molecules, including nucleic acids, proteins, and lipids. Recently, the role of extracellular vesicle signaling in cancer progression has become a topic of significant interest. Methods to detect and target exosomes and microvesicles are needed to realize applications of extracellular vesicles as biomarkers and, perhaps, therapeutic targets. Detection of exosomes and microvesicles is a complex problem as they are both submicroscopic and of heterogeneous cellular origins. In this Minireview, we highlight the basic biology of extracellular vesicles, and address available biochemical and biophysical detection methods. Detectible characteristics described here include lipid and protein composition, and physical properties such as the vesicle membrane shape and diffusion coefficient. In particular, we propose that detection of exosome and microvesicle membrane curvature with lipid chemical probes that sense membrane shape is a distinctly promising method for identifying and targeting these vesicles.
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Extracellular Vesicles
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Cells release multiple, distinct forms of extracellular vesicles including structures known as microvesicles, which are known to alter the extracellular environment. Despite growing understanding of microvesicle biogenesis, function and contents, mechanisms regulating cargo delivery and enrichment remain largely unknown. Here we demonstrate that in amoeboid-like invasive tumour cell lines, the v-SNARE, VAMP3, regulates delivery of microvesicle cargo such as the membrane-type 1 matrix metalloprotease (MT1-MMP) to shedding microvesicles. MT1-MMP delivery to nascent microvesicles depends on the association of VAMP3 with the tetraspanin CD9 and facilitates the maintenance of amoeboid cell invasion. VAMP3-shRNA expression depletes shed vesicles of MT1-MMP and decreases cell invasiveness when embedded in cross-linked collagen matrices. Finally, we describe functionally similar microvesicles isolated from bodily fluids of ovarian cancer patients. Together these studies demonstrate the importance of microvesicle cargo sorting in matrix degradation and disease progression. Cells shed various types of vesicles differing in size and content. Here the authors show that cancer cells utilize VAMP3-mediated traffic to deliver MT1-MMP to surface microvesicles and facilitate amoeboid-like cell invasion, with VAMP3-containing vesicles also found in body fluids of cancer patients.
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Microvesicle biogenesis is a highly regulated process. Aberrant release of microvesicles from cancer cells have been associated with their invasiveness and prognosis. However, the mechanism of aberrant release remains poorly understood. Herein, we found that hepatocellular carcinoma cells shed more microvesicles than normal hepatocytes and miR-200a were shown to inhibit the release of microvesicles in hepatocellular carcinoma cells. Then, we confirmed that miR-200a might target Gelsolin and change cytoskeleton to regulate microvesicles secretion. Further miR-200a may inhibit the proliferation of adjacent cells by inhibiting the release of microvesicles. Collectively, our findings indicate that miR-200a regulated the microvesicle biogenesis involved in the hepatocellular carcinoma progression.
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Objective To evaluate the expression of several CD cell markers and investigate their significance in hepatocellular carcinoma cell lines with different metastatic potential. Methods Three human hepatocellular cell lines,HepG2,MHCC-97H and SK-HEP-1 were cultivated by routine method.Single cell suspension was achieved by digestion with trypsin.After CD133,CD90,CD44, CD34 and CD24 antibody labeling,cells were analyzed by flow cytometry. Results CD133+ cells represented only a very small subpopulation in three hepatocellular carcinoma cell lines(0.1%~0.4%).In addition,CD90+,CD44+ and CD90+CD44+cells were detected in the three cell lines and the percentage increased with the development of metastatic potential(P﹤0.05).The expression rates of CD44+CD24+ cells in HepG2,MHCC-97H and SK-HEP-1 were 0.05%,10.3% and 0.3%,respectively(P﹤0.05).Furthermore,no CD34+ or CD133+CD90+ expression was observed in the three cell lines. Conclusion CD90 and CD44 are significantly correlated with the metastatic potential of hepatocellular carcinoma cells and may be prospective markers for hepatocellular carcinoma stem cells.
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