Using 3D Invasion properties of RCC Cell Lines In Vitro to predict their Metastatic Potential In Vivo

Using 3D Invasion properties of RCC Cell Lines In Vitro to predict their Metastatic Potential In Vivo | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Using 3D Invasion properties of RCC Cell Lines In Vitro to predict their Metastatic Potential In Vivo Odile Filhol, Beatrice Cesana, Laurie Nemoz-Billet, Valentin Azemard, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6812883/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 27 Feb, 2026 Read the published version in Cell Death Discovery → Version 1 posted 11 You are reading this latest preprint version Abstract Renal cell carcinoma (RCC) exhibits significant heterogeneity, making it challenging to predict tumor aggressiveness and therapeutic response. To improve prognostic accuracy and develop tailored treatment strategies, it is crucial to mimic both cancer cells and their microenvironment in vitro. Using a combination of in vitro and in vivo models, we investigated the invasive properties of three RCC cell lines, RCC10, RCC7 and 786-O, that displayed distinct signaling profiles, combining EMT characteristics and upregulation of key metastatic markers. Our findings revealed that RCC7 and 786-O exhibited greater metastatic potential than RCC10, as demonstrated by increased extravasation in zebrafish embryos and higher lung metastases in the chorioallantoic membrane (CAM) and mice models. Comparative pathway analysis indicated that RCC7 displays partial epithelial-mesenchymal transition (pEMT) characteristics and upregulates key metastatic markers. Furthermore, our 3D spheroid invasion model as well as our patient-derived RCC tumoroid system predicted accurately their metastatic behavior, closely mirroring their aggressiveness in vivo. Thus, these 3D models might be predictive of tumor outcome, underscoring their utility as reliable predictive tools for RCC progression and therapeutic response. Biological sciences/Cancer/Urological cancer/Renal cancer/Renal cell carcinoma Health sciences/Diseases/Cancer/Metastasis renal cell carcinoma 3D models patient-derived tumoroids invasion assays zebrafish embryos Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Full Text Additional Declarations (Not answered) Supplementary Files SupplementalMaterial.pdf supplemental material SupplentalMaterial2.pdf Western blots RCC7.avi RCC7 invasion RCC10.avi RCC10 invasion 786O.avi 786-O Invasion Cite Share Download PDF Status: Published Journal Publication published 27 Feb, 2026 Read the published version in Cell Death Discovery → Version 1 posted Unknown event 06 Nov, 2025 Editorial decision: Reject after peer review 11 Jul, 2025 Review # 1 received at journal 01 Jul, 2025 Review # 2 received at journal 25 Jun, 2025 Reviewer # 2 agreed at journal 14 Jun, 2025 Reviewer # 1 agreed at journal 13 Jun, 2025 Reviewers invited by journal 13 Jun, 2025 Submission checks completed at journal 05 Jun, 2025 First submitted to journal 04 Jun, 2025 Unknown event 04 Jun, 2025 Editor assigned by journal 03 Jun, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6812883","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":470941879,"identity":"04abb92a-b155-4837-bd1b-dee82c214f0c","order_by":0,"name":"Odile Filhol","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCElEQVRIie3QsUrDQBzH8V84uCxHs6a01Ff4S4YuYl8l5aAubi4BC54InQ5cfQxXtwuBdOkDRBxM8AU6ZqhirnZQyGV2uO/0T7gPuX8An+9ftrw3wEyMYgTGPhMQ1HYIlZOo7mQieAycCGNkB2FcJLUnE/wmdnaTKJTK7EFTPnkwBuuLs3n4WGZtdgkx6Tdj3aj8CST4tEwNytX5iy74q95JiFHaS6haqkLg0O1yTcWnKlKqJH8LNgwL0X+xIznYr3TEBJa8f/Cb4OsOYojgD6kYZ93gJMddNFmy+tnleSeTsS63wkWi8Kqp24wWUSzz2v4x2ubNvl3fzlzkdL2ed4PA5/P5fMN9A6ScWN6TXSkVAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0003-1964-7958","institution":"University of Grenoble Alpes, Inserm, CEA UMR1292","correspondingAuthor":true,"prefix":"","firstName":"Odile","middleName":"","lastName":"Filhol","suffix":""},{"id":470941880,"identity":"9d0a2fed-eba5-42fd-ae31-875bb6a110c3","order_by":1,"name":"Beatrice Cesana","email":"","orcid":"","institution":"University of Grenoble Alpes, Inserm, CEA UMR1292","correspondingAuthor":false,"prefix":"","firstName":"Beatrice","middleName":"","lastName":"Cesana","suffix":""},{"id":470941881,"identity":"b75bc3e5-c12f-4c82-8457-c6cd949f2125","order_by":2,"name":"Laurie Nemoz-Billet","email":"","orcid":"","institution":"Inserm UMR_S1109","correspondingAuthor":false,"prefix":"","firstName":"Laurie","middleName":"","lastName":"Nemoz-Billet","suffix":""},{"id":470941882,"identity":"255103ab-5167-4d22-a68c-ddb2e304664d","order_by":3,"name":"Valentin Azemard","email":"","orcid":"","institution":"University of Grenoble Alpes, Inserm, CEA UMR1292","correspondingAuthor":false,"prefix":"","firstName":"Valentin","middleName":"","lastName":"Azemard","suffix":""},{"id":470941883,"identity":"c74a0367-5aef-4499-b42b-9ebd1a995115","order_by":4,"name":"Catherine Pillet","email":"","orcid":"","institution":"University of Grenoble Alpes, Inserm, CEA UMR1292","correspondingAuthor":false,"prefix":"","firstName":"Catherine","middleName":"","lastName":"Pillet","suffix":""},{"id":470941884,"identity":"81d7d23c-c9f3-4d82-93ae-bc6d876fb797","order_by":5,"name":"Estelle Bigot","email":"","orcid":"","institution":"University of Grenoble Alpes, Inserm, CEA UMR1292","correspondingAuthor":false,"prefix":"","firstName":"Estelle","middleName":"","lastName":"Bigot","suffix":""},{"id":470941885,"identity":"124c14ad-8d4e-4de7-96da-3f498fbf66c7","order_by":6,"name":"Nicolas Chaumontel","email":"","orcid":"","institution":"University of Grenoble Alpes, Inserm, CEA UMR1292","correspondingAuthor":false,"prefix":"","firstName":"Nicolas","middleName":"","lastName":"Chaumontel","suffix":""},{"id":470941886,"identity":"67d82d38-19bf-4095-9843-80c7cbf6f193","order_by":7,"name":"Jean-Luc Descotes","email":"","orcid":"","institution":"Centre hospitalier universitaire Grenoble Alpes","correspondingAuthor":false,"prefix":"","firstName":"Jean-Luc","middleName":"","lastName":"Descotes","suffix":""},{"id":470941887,"identity":"b4bac561-6c49-460b-b4ff-156dfaf4e41c","order_by":8,"name":"Naël Osmani","email":"","orcid":"","institution":"Inserm UMR_S1109","correspondingAuthor":false,"prefix":"","firstName":"Naël","middleName":"","lastName":"Osmani","suffix":""},{"id":470941888,"identity":"8de74a0c-0433-43b9-bd5c-d220d640a8e0","order_by":9,"name":"Jacky Goetz","email":"","orcid":"","institution":"Inserm UMR_S1109","correspondingAuthor":false,"prefix":"","firstName":"Jacky","middleName":"","lastName":"Goetz","suffix":""},{"id":470941889,"identity":"3004b02b-5fec-434e-846d-7b12db3f7d6e","order_by":10,"name":"Claude Cochet","email":"","orcid":"","institution":"Institut National de la Sant� et de la Recherche M�dicale","correspondingAuthor":false,"prefix":"","firstName":"Claude","middleName":"","lastName":"Cochet","suffix":""}],"badges":[],"createdAt":"2025-06-03 15:26:37","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6812883/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6812883/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41420-026-02966-7","type":"published","date":"2026-02-27T05:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":85266795,"identity":"4baa6cfd-e814-4f03-b0d7-08bf0e33804f","added_by":"auto","created_at":"2025-06-24 05:41:05","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":636655,"visible":true,"origin":"","legend":"\u003cp\u003e3D spheroid invasion assay. (A) Representative images of RCC7, 786-O and RCC10 spheroids cultured in Collagen I and \u0026nbsp;\u0026nbsp;Fibronectin hydrogel visualized after 7 days with Live and Dead \u0026nbsp;\u0026nbsp;staining (Calcein staining living cells in green, Hoechst 33342 \u0026nbsp;staining nuclei in blue and Ethidium Homodimer I staining \u0026nbsp;dead cells in red). Scale bar = 300 µm. (B) Maximum invasion \u0026nbsp;length of RCC cell lines invading through hydrogel after 7 days \u0026nbsp;of culture. (C) Average invasion area of ccRCC cell lines \u0026nbsp;invading through hydrogel after 7 days of culture. (D) Number \u0026nbsp;of cells invading the hydrogel after 7 days of culture. (E) \u0026nbsp;Invasion speed of each cell line. (F) Invasion speed of RCC7, \u0026nbsp;786-O and RCC10 spheroids embedded in hydrogel measured \u0026nbsp;during 5 days. (G) Representative rose plot of the invasion of \u0026nbsp;cells through hydrogel. Data show mean ± SEM, RCC7 n = 10, \u0026nbsp;786-O n = 16, RCC10 n = 15, ACHN n = 12. Significance was \u0026nbsp;assessed using a one-way ANOVA with Tukey’s multiple \u0026nbsp;comparison test comparing each cell line to every other cell \u0026nbsp;line. (*p\u0026lt;0.05;**** p\u0026lt;0.0001)\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1/269b9b3c56b4386d49307f7b.png"},{"id":85266796,"identity":"3edfc5f4-b0fd-4c6c-a79b-e57d9ffb6cfe","added_by":"auto","created_at":"2025-06-24 05:41:05","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":2302852,"visible":true,"origin":"","legend":"\u003cp\u003eRCC cells have different tumor growth and metastatic \u0026nbsp;potential in the Chick embryo chorio-allantoic membrane \u0026nbsp;(CAM) model. (A) Scheme of the experimental approach of \u0026nbsp;RCC cells injection into the Chorio-Allantoic Membrane of \u0026nbsp;chick embryos. (B) Representative images of tumor derived \u0026nbsp;from RCC10, 786-O and RCC7 cells in ovo at E16. For RCC7 \u0026nbsp;tumors, the important accumulation of blood prevents to \u0026nbsp;distinctly locate the tumor. (C) Illustrative pictures of RCC10, \u0026nbsp;786-O and RCC7-induced tumors (post-collection). (D) Mean \u0026nbsp;tumor weight (mg) measured in the different experimental \u0026nbsp;groups at the end of the study. (E) Number of embryos that \u0026nbsp;showed a positive signal in the analysis of the metastatic \u0026nbsp;invasion, measured by qPCR for human specific Alu sequences \u0026nbsp;in the lower CAM and in the lungs. Data are shown as mean \u0026nbsp;with SEM with n = 8 animals/group and n = 3 for the control. \u0026nbsp;Significance was assessed using a one-way ANOVA with \u0026nbsp;Tukey’s multiple comparison test comparing each cell line to \u0026nbsp;every other cell line. (**** p\u0026lt;0.001).\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1/e3c2a87ceb1933845204821f.png"},{"id":85266799,"identity":"b5e6d68d-e5e5-4177-a4fc-269440fcedc7","added_by":"auto","created_at":"2025-06-24 05:41:06","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":2814327,"visible":true,"origin":"","legend":"\u003cp\u003eAdhesion and extravasation capacities of RCC cells in \u0026nbsp;zebrafish in vivo model. (A) Scheme of the experimental \u0026nbsp;approach using 2 dpf Tg(Fli1:EGFP) zebrafish embryos as a \u0026nbsp;metastatic cascade model. Right panel: blood flow in the \u0026nbsp;caudal plexus regions: dorsal aorta (DA), arterio-venous \u0026nbsp;junction (AVJ), caudal veins (CVs). The adhesion pattern of \u0026nbsp;RCC7, 786-O or RCC10 cells was imaged 3, 24 and 48 hours \u0026nbsp;post-injection (hpi). (B) Total number of arrested cells at 3 hpi. \u0026nbsp;The graph shows the mean ± SEM of 2 independent \u0026nbsp;experiments (RCC10 n = 19; 786-O = 27; RCC7 n = 28). (C) Total \u0026nbsp;number of arrested cells at 24 hpi (D) Representative images \u0026nbsp;of cells arrested at 24 hpi. (E) The heatmaps show \u0026nbsp;quantification of the number and location of arrested RCC cells \u0026nbsp;at 24 hpi in the caudal plexus. (F) Ratio of cells adhered at 24 \u0026nbsp;hpi over the total number of cells and their localization (see A) \u0026nbsp;of cells adhered at 24 hpi were measured. The graphs shows \u0026nbsp;the mean ± SEM of 2 independent experiments (RCC10 n = 13; \u0026nbsp;786-O = 16; RCC7 n = 20). (G) Total number of arrested cells at \u0026nbsp;48 hpi. The graph shows the mean ± SEM (RCC10 n = 12; 786- \u0026nbsp;O n = 13; RCC7 n = 11). (H) Representative images (left) and its \u0026nbsp;orthoslice (right) of extravascular cells at 48 hpi. (I) The \u0026nbsp;heatmaps show quantification of the number and location of \u0026nbsp;extravascular RCC cells at 48 hpi in the caudal plexus. (J) Ratio \u0026nbsp;of cells extravasated over the total number of cells was \u0026nbsp;measured at 48 hpi Significance was assessed using a one-way \u0026nbsp;ANOVA with Tukey’s multiple comparison test comparing each \u0026nbsp;cell line to every other cell line. (*p\u0026lt;0.05; ** p\u0026lt;0.01; *** \u0026nbsp;p\u0026lt;0.001; **** p\u0026lt;0.0001).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1/3e78928fb0fa1b93f7d86a29.png"},{"id":85266804,"identity":"033a3bfd-e3a0-4b5d-b4a9-3425c18e6abb","added_by":"auto","created_at":"2025-06-24 05:41:06","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":2471395,"visible":true,"origin":"","legend":"\u003cp\u003eMetastatic Potential of RCC Cells in Murine Lungs. (A) \u0026nbsp;Scheme of the experimental approach of retro-orbital \u0026nbsp;injection of luciferase-transduced RCC cells into BALB/c Nude \u0026nbsp;female mice. Lungs, liver, femoral bone and kidney were \u0026nbsp;specifically screened for metastasis. (B) Number of mice that \u0026nbsp;developed lung metastases for each cell lines. (C) Time of \u0026nbsp;metastasis onset after retro-orbital injection. (D) Average \u0026nbsp;radiance intensity measured after luciferin injection (200µl, 15 \u0026nbsp;mg/ml) with IVIS® Spectrum. (E) Representative tissue \u0026nbsp;sections stained with H\u0026amp;E, obtained from lungs of mice after \u0026nbsp;retro-orbital injection of RCC cells. Scale bar = 100 µm. Data \u0026nbsp;are shown as mean with SEM with n=3 animals/group. \u0026nbsp;Significance was assessed using a one-way ANOVA with \u0026nbsp;Tukey’s multiple comparison test comparing each cell line to \u0026nbsp;every other cell line. (*p\u0026lt;0.05; ** p\u0026lt;0.01).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1/6c9eaed15ec404fd3157a357.png"},{"id":85266803,"identity":"3d44cd2a-8d8e-4b2f-92c1-fa6ba475d058","added_by":"auto","created_at":"2025-06-24 05:41:06","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1544351,"visible":true,"origin":"","legend":"\u003cp\u003e3D human renal tumoroid invasion assay. (A) \u0026nbsp;Representative images of MC-156 and PD-157 tumoroids \u0026nbsp;cultured in Collagen I and Fibronectin hydrogel, at 0, 3 and 7 \u0026nbsp;days of culture. (Scale bar = 100 µm). (B) Maximum invasion \u0026nbsp;length of RCC cells invading through hydrogel after 7 days of \u0026nbsp;culture. (C) Average invasion area of RCC cells invading \u0026nbsp;through hydrogel after 7 days of culture. (D) Invasion speed of \u0026nbsp;RCC7, 786-O and RCC10 spheroids embedded in hydrogel \u0026nbsp;measured during 5 days. (E) Representative rose plot of the \u0026nbsp;invasion of tumoroid cells through hydrogel. Data show mean \u0026nbsp;± SEM, MC-156 n = 12, PD-157 n = 12. (F) Immunostaining of \u0026nbsp;MC-156 invading cells (CAIX antibody staining renal cells in \u0026nbsp;red, phalloidin staining F-actin in green and Hoechst staining \u0026nbsp;nuclei in blue). Significance was assessed using a one-way \u0026nbsp;ANOVA with Tukey’s multiple comparison test comparing each \u0026nbsp;cell line to every other cell line. **** p\u0026lt;0.0001).\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1/af668387fc4b215049615536.png"},{"id":105618277,"identity":"ea2aed61-8588-4cd0-bb65-f7ef8b4d783f","added_by":"auto","created_at":"2026-03-28 07:12:59","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2081607,"visible":true,"origin":"","legend":"Article File","description":"","filename":"Cesanaetal.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1_covered_cb60ee3a-8fde-40eb-9148-30eb39d5e837.pdf"},{"id":85266798,"identity":"46fa2724-51db-4df4-aded-eaedd427fe71","added_by":"auto","created_at":"2025-06-24 05:41:06","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":930455,"visible":true,"origin":"","legend":"supplemental material","description":"","filename":"SupplementalMaterial.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1/e56693bd19c1b256344a5365.pdf"},{"id":85266801,"identity":"b78b7300-13de-4949-880c-3dfd6fa52de2","added_by":"auto","created_at":"2025-06-24 05:41:06","extension":"pdf","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":604082,"visible":true,"origin":"","legend":"Western blots","description":"","filename":"SupplentalMaterial2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1/1e543d2aa9e05b87856b9ebf.pdf"},{"id":85266807,"identity":"3fbd52fd-8eb1-47ae-b61c-8ceb5d6ece27","added_by":"auto","created_at":"2025-06-24 05:41:07","extension":"avi","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":76445176,"visible":true,"origin":"","legend":"RCC7 invasion","description":"","filename":"RCC7.avi","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1/25067445f51359794f9bb2bb.avi"},{"id":85266809,"identity":"b0afd280-df37-4002-964f-b28b40f1f551","added_by":"auto","created_at":"2025-06-24 05:41:08","extension":"avi","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":95812702,"visible":true,"origin":"","legend":"RCC10 invasion","description":"","filename":"RCC10.avi","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1/6a6122901e9e40f55594fc2d.avi"},{"id":85266808,"identity":"bb1e27e5-82d0-4caf-8a79-ea1698ba24c2","added_by":"auto","created_at":"2025-06-24 05:41:07","extension":"avi","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":99626492,"visible":true,"origin":"","legend":"786-O Invasion","description":"","filename":"786O.avi","url":"https://assets-eu.researchsquare.com/files/rs-6812883/v1/61604d112876410ef4f09a80.avi"}],"financialInterests":"(Not answered)","formattedTitle":"\u003cp\u003eUsing 3D Invasion properties of RCC Cell Lines \u003cem\u003eIn Vitro\u003c/em\u003e to predict their Metastatic Potential\u003cem\u003e In Vivo\u003c/em\u003e\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"cell-death-discovery","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"cddiscovery","sideBox":"Learn more about [Cell Death Discovery](http://www.nature.com/cddiscovery/)","snPcode":"41420","submissionUrl":"https://mts-cddiscovery.nature.com/","title":"Cell Death Discovery","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"renal cell carcinoma, 3D models, patient-derived tumoroids, invasion assays, zebrafish embryos","lastPublishedDoi":"10.21203/rs.3.rs-6812883/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6812883/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Renal cell carcinoma (RCC) exhibits significant heterogeneity, making it challenging to predict tumor aggressiveness and therapeutic response. To improve prognostic accuracy and develop tailored treatment strategies, it is crucial to mimic both cancer cells and their microenvironment in vitro. Using a combination of in vitro and in vivo models, we investigated the invasive properties of three RCC cell lines, RCC10, RCC7 and 786-O, that displayed distinct signaling profiles, combining EMT characteristics and upregulation of key metastatic markers. Our findings revealed that RCC7 and 786-O exhibited greater metastatic potential than RCC10, as demonstrated by increased extravasation in zebrafish embryos and higher lung metastases in the chorioallantoic membrane (CAM) and mice models. Comparative pathway analysis indicated that RCC7 displays partial epithelial-mesenchymal transition (pEMT) characteristics and upregulates key metastatic markers. Furthermore, our 3D spheroid invasion model as well as our patient-derived RCC tumoroid system predicted accurately their metastatic behavior, closely mirroring their aggressiveness in vivo. Thus, these 3D models might be predictive of tumor outcome, underscoring their utility as reliable predictive tools for RCC progression and therapeutic response.","manuscriptTitle":"Using 3D Invasion properties of RCC Cell Lines In Vitro to predict their Metastatic Potential In Vivo","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-24 05:41:01","doi":"10.21203/rs.3.rs-6812883/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"transferred","content":"Cell Death Discovery","date":"2025-11-06T11:36:52+00:00","index":"","fulltext":""},{"type":"decision","content":"Reject after peer review","date":"2025-07-11T16:40:31+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"This content is not available.","date":"2025-07-01T10:06:21+00:00","index":1,"fulltext":"This content is not available."},{"type":"editorInvitedReview","content":"This content is not available.","date":"2025-06-25T09:41:43+00:00","index":2,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2025-06-14T08:15:53+00:00","index":2,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2025-06-13T16:09:00+00:00","index":1,"fulltext":"This content is not available."},{"type":"reviewersInvited","content":"","date":"2025-06-13T13:34:59+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-06-05T11:36:17+00:00","index":"","fulltext":""},{"type":"submitted","content":"Cell Death \u0026 Disease","date":"2025-06-04T13:21:07+00:00","index":"","fulltext":""},{"type":"checksFailed","content":"","date":"2025-06-04T11:07:43+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-06-03T15:23:34+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"cell-death-discovery","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"cddiscovery","sideBox":"Learn more about [Cell Death Discovery](http://www.nature.com/cddiscovery/)","snPcode":"41420","submissionUrl":"https://mts-cddiscovery.nature.com/","title":"Cell Death Discovery","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"c4f10b27-99ae-45bc-a919-c4d151c20197","owner":[],"postedDate":"June 24th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":50201730,"name":"Biological sciences/Cancer/Urological cancer/Renal cancer/Renal cell carcinoma"},{"id":50201731,"name":"Health sciences/Diseases/Cancer/Metastasis"}],"tags":[],"updatedAt":"2026-03-28T07:12:47+00:00","versionOfRecord":{"articleIdentity":"rs-6812883","link":"https://doi.org/10.1038/s41420-026-02966-7","journal":{"identity":"cell-death-discovery","isVorOnly":false,"title":"Cell Death Discovery"},"publishedOn":"2026-02-27 05:00:00","publishedOnDateReadable":"February 27th, 2026"},"versionCreatedAt":"2025-06-24 05:41:01","video":"","vorDoi":"10.1038/s41420-026-02966-7","vorDoiUrl":"https://doi.org/10.1038/s41420-026-02966-7","workflowStages":[]},"version":"v1","identity":"rs-6812883","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6812883","identity":"rs-6812883","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}