En bloc release of MVB-like small extracellular vesicle clusters by colorectal carcinoma cells
Small extracellular vesicles (EVs) are membrane enclosed structures that are usually released from cells upon exocytosis of multivesicular bodies (MVBs) as a collection of separate, free EVs. In this study, we analysed paraffin embedded sections of archived human colorectal cancer samples. We studie...
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| Veröffentlicht in: | Journal of extracellular vesicles 2019-12, Vol.8 (1), p.1596668-n/a |
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| creator | Valcz, Gábor Buzás, Edit I. Kittel, Ágnes Krenács, Tibor Visnovitz, Tamás Spisák, Sándor Török, György Homolya, László Zsigrai, Sára Kiszler, Gábor Antalffy, Géza Pálóczi, Krisztina Szállási, Zoltán Szabó, Vanessza Sebestyén, Anna Solymosi, Norbert Kalmár, Alexandra Dede, Kristóf Lőrincz, Péter Tulassay, Zsolt Igaz, Péter Molnár, Béla |
| description | Small extracellular vesicles (EVs) are membrane enclosed structures that are usually released from cells upon exocytosis of multivesicular bodies (MVBs) as a collection of separate, free EVs. In this study, we analysed paraffin embedded sections of archived human colorectal cancer samples. We studied 3D reconstructions of confocal microscopic images complemented by HyVolution and STED imaging. Unexpectedly, we found evidence that large, MVB-like aggregates of ALIX/CD63 positive EV clusters were released en bloc by migrating tumour cells. These structures were often captured with partial or complete extra-cytoplasmic localization at the interface of the plasma membrane of the tumour cell and the stroma. Their diameter ranged between 0.62 and 1.94 μm (mean±S.D.: 1.17 ± 0.34 μm). High-resolution 3D reconstruction showed that these extracellular MVB-like EV clusters were composed of distinguishable internal particles of small EV size (mean±S.D.: 128.96 ± 16.73 nm). In vitro, HT29 colorectal cancer cells also showed the release of similar structures as confirmed by immunohistochemistry and immune electron microscopy. Our results provide evidence for an en bloc transmission of MVB-like EV clusters through the plasma membrane. Immunofluorescent-based detection of the MVB like small EV clusters in archived pathological samples may represent a novel and unique opportunity which enables analysis of EV release in situ in human tissues. |
| doi_str_mv | 10.1080/20013078.2019.1596668 |
| format | Article |
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In this study, we analysed paraffin embedded sections of archived human colorectal cancer samples. We studied 3D reconstructions of confocal microscopic images complemented by HyVolution and STED imaging. Unexpectedly, we found evidence that large, MVB-like aggregates of ALIX/CD63 positive EV clusters were released en bloc by migrating tumour cells. These structures were often captured with partial or complete extra-cytoplasmic localization at the interface of the plasma membrane of the tumour cell and the stroma. Their diameter ranged between 0.62 and 1.94 μm (mean±S.D.: 1.17 ± 0.34 μm). High-resolution 3D reconstruction showed that these extracellular MVB-like EV clusters were composed of distinguishable internal particles of small EV size (mean±S.D.: 128.96 ± 16.73 nm). In vitro, HT29 colorectal cancer cells also showed the release of similar structures as confirmed by immunohistochemistry and immune electron microscopy. Our results provide evidence for an en bloc transmission of MVB-like EV clusters through the plasma membrane. Immunofluorescent-based detection of the MVB like small EV clusters in archived pathological samples may represent a novel and unique opportunity which enables analysis of EV release in situ in human tissues.</description><identifier>ISSN: 2001-3078</identifier><identifier>EISSN: 2001-3078</identifier><identifier>DOI: 10.1080/20013078.2019.1596668</identifier><identifier>PMID: 31007874</identifier><language>eng</language><publisher>Sweden: Taylor & Francis</publisher><subject>Antibodies ; Antigens ; Binding sites ; Cancer ; CD63 antigen ; Cell culture ; Colorectal carcinoma ; Cytokeratin ; Electron microscopy ; Exocytosis ; Extracellular vesicles ; Immunohistochemistry ; Laboratories ; Localization ; Membrane vesicles ; Microscopy ; migrating cancer cells ; MVB ; Proteins ; Stroma ; Tumors</subject><ispartof>Journal of extracellular vesicles, 2019-12, Vol.8 (1), p.1596668-n/a</ispartof><rights>2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles. 2019</rights><rights>2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles.</rights><rights>2019. This work is published under http://creativecommons.org/licenses/by-nc/4.0/ (the "License"). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles. 2019 The Author(s)</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c6177-96153ca5d66271f91ef20675a52cfddd55111225c4e5604437604318cd3316023</citedby><cites>FETCH-LOGICAL-c6177-96153ca5d66271f91ef20675a52cfddd55111225c4e5604437604318cd3316023</cites><orcidid>0000-0003-2192-554X ; 0000-0001-7374-667X ; 0000-0003-1639-8140 ; 0000-0002-3744-206X ; 0000-0001-9164-065X ; 0000-0002-3824-1534 ; 0000-0001-5147-1273 ; 0000-0003-2452-6640</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6461071/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6461071/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,724,777,781,861,882,1412,2096,11543,27483,27905,27906,45555,45556,46033,46457,53772,53774,59122,59123</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31007874$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Valcz, Gábor</creatorcontrib><creatorcontrib>Buzás, Edit I.</creatorcontrib><creatorcontrib>Kittel, Ágnes</creatorcontrib><creatorcontrib>Krenács, Tibor</creatorcontrib><creatorcontrib>Visnovitz, Tamás</creatorcontrib><creatorcontrib>Spisák, Sándor</creatorcontrib><creatorcontrib>Török, György</creatorcontrib><creatorcontrib>Homolya, László</creatorcontrib><creatorcontrib>Zsigrai, Sára</creatorcontrib><creatorcontrib>Kiszler, Gábor</creatorcontrib><creatorcontrib>Antalffy, Géza</creatorcontrib><creatorcontrib>Pálóczi, Krisztina</creatorcontrib><creatorcontrib>Szállási, Zoltán</creatorcontrib><creatorcontrib>Szabó, Vanessza</creatorcontrib><creatorcontrib>Sebestyén, Anna</creatorcontrib><creatorcontrib>Solymosi, Norbert</creatorcontrib><creatorcontrib>Kalmár, Alexandra</creatorcontrib><creatorcontrib>Dede, Kristóf</creatorcontrib><creatorcontrib>Lőrincz, Péter</creatorcontrib><creatorcontrib>Tulassay, Zsolt</creatorcontrib><creatorcontrib>Igaz, Péter</creatorcontrib><creatorcontrib>Molnár, Béla</creatorcontrib><title>En bloc release of MVB-like small extracellular vesicle clusters by colorectal carcinoma cells</title><title>Journal of extracellular vesicles</title><addtitle>J Extracell Vesicles</addtitle><description>Small extracellular vesicles (EVs) are membrane enclosed structures that are usually released from cells upon exocytosis of multivesicular bodies (MVBs) as a collection of separate, free EVs. 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Our results provide evidence for an en bloc transmission of MVB-like EV clusters through the plasma membrane. Immunofluorescent-based detection of the MVB like small EV clusters in archived pathological samples may represent a novel and unique opportunity which enables analysis of EV release in situ in human tissues.</description><subject>Antibodies</subject><subject>Antigens</subject><subject>Binding sites</subject><subject>Cancer</subject><subject>CD63 antigen</subject><subject>Cell culture</subject><subject>Colorectal carcinoma</subject><subject>Cytokeratin</subject><subject>Electron microscopy</subject><subject>Exocytosis</subject><subject>Extracellular vesicles</subject><subject>Immunohistochemistry</subject><subject>Laboratories</subject><subject>Localization</subject><subject>Membrane vesicles</subject><subject>Microscopy</subject><subject>migrating cancer cells</subject><subject>MVB</subject><subject>Proteins</subject><subject>Stroma</subject><subject>Tumors</subject><issn>2001-3078</issn><issn>2001-3078</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>0YH</sourceid><sourceid>24P</sourceid><sourceid>WIN</sourceid><sourceid>DOA</sourceid><recordid>eNqNUk1v1DAQjRCIVqU_AWSJC5ddPHbsJBcErbZQVMQFesRy7Enx4sStnbTsv8dht6XlgPDBtubjzcybVxTPgS6B1vQ1oxQ4reolo9AsQTRSyvpRsT_bF7Pj8b3_XnGY0prm05Qg6uZpsceBZkdV7hffVgNpfTAkokedkISOfDo_Wnj3A0nqtfcEf45RG_R-8jqSa0zOeCTGT2nEmEi7ISb4ENGM2hOjo3FD6DWZM9Kz4kmnfcLD3XtQfD1ZfTn-sDj7_P70-N3ZwkioqkUjQXCjhZWSVdA1gB2jshJaMNNZa4UAAMaEKVFIWpa8yjeH2ljOQVLGD4rTLa4Neq0uo-t13KignfptCPFC6TjOjavMg6xNzVpetqWlXaNpy2ydabWYa1cZ680W63Jqe7QGhzy_fwD60DO47-oiXCtZSqAVZIBXO4AYriZMo-pdmunQA4YpKcYgTykbOdd6-VfoOkxxyFQpThvGy7wvnqPENsrEkFLE7q4ZoGoWhLoVhJoFoXaCyHkv7k9yl3W7_j-j3jiPm_9DVR9X5-zoJNNYz_2_3QK4oQux1zcheqtGvcmC6KIejEtztX81-QtCCdQh</recordid><startdate>201912</startdate><enddate>201912</enddate><creator>Valcz, Gábor</creator><creator>Buzás, Edit I.</creator><creator>Kittel, Ágnes</creator><creator>Krenács, Tibor</creator><creator>Visnovitz, Tamás</creator><creator>Spisák, Sándor</creator><creator>Török, György</creator><creator>Homolya, László</creator><creator>Zsigrai, Sára</creator><creator>Kiszler, Gábor</creator><creator>Antalffy, Géza</creator><creator>Pálóczi, Krisztina</creator><creator>Szállási, Zoltán</creator><creator>Szabó, Vanessza</creator><creator>Sebestyén, Anna</creator><creator>Solymosi, Norbert</creator><creator>Kalmár, Alexandra</creator><creator>Dede, Kristóf</creator><creator>Lőrincz, Péter</creator><creator>Tulassay, Zsolt</creator><creator>Igaz, Péter</creator><creator>Molnár, Béla</creator><general>Taylor & Francis</general><general>John Wiley & Sons, Inc</general><general>Wiley</general><scope>0YH</scope><scope>24P</scope><scope>WIN</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QP</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0003-2192-554X</orcidid><orcidid>https://orcid.org/0000-0001-7374-667X</orcidid><orcidid>https://orcid.org/0000-0003-1639-8140</orcidid><orcidid>https://orcid.org/0000-0002-3744-206X</orcidid><orcidid>https://orcid.org/0000-0001-9164-065X</orcidid><orcidid>https://orcid.org/0000-0002-3824-1534</orcidid><orcidid>https://orcid.org/0000-0001-5147-1273</orcidid><orcidid>https://orcid.org/0000-0003-2452-6640</orcidid></search><sort><creationdate>201912</creationdate><title>En bloc release of MVB-like small extracellular vesicle clusters by colorectal carcinoma cells</title><author>Valcz, Gábor ; 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| ispartof | Journal of extracellular vesicles, 2019-12, Vol.8 (1), p.1596668-n/a |
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| source | Taylor & Francis Open Access; DOAJ Directory of Open Access Journals; Wiley Online Library Journals Frontfile Complete; Co-Action Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Wiley Online Library Open Access; PubMed Central |
| subjects | Antibodies Antigens Binding sites Cancer CD63 antigen Cell culture Colorectal carcinoma Cytokeratin Electron microscopy Exocytosis Extracellular vesicles Immunohistochemistry Laboratories Localization Membrane vesicles Microscopy migrating cancer cells MVB Proteins Stroma Tumors |
| title | En bloc release of MVB-like small extracellular vesicle clusters by colorectal carcinoma cells |
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