UGGT1 mediates N-glycosylation of LGALS3BP to induce angiogenesis in liver cancer via the NOTCH signaling pathway

UGGT1 mediates N-glycosylation of LGALS3BP to induce angiogenesis in liver cancer via the NOTCH signaling pathway | 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 Research Article UGGT1 mediates N-glycosylation of LGALS3BP to induce angiogenesis in liver cancer via the NOTCH signaling pathway Benzhen He, Liang Wang, Jiadong Xia, Xinggang Xu, Jianguo Xu, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8052281/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Background Liver cancer (LC) is one of the leading causes of cancer-related deaths worldwide. Its complex vascular regulatory network and drug resistance severely limit clinical efficacy. Although LGALS3BP is highly expressed in various malignant tumors, its biological function and molecular regulatory mechanisms in LC angiogenesis remain to be elucidated. Methods Bioinformatics analysis was used to examine the expression of LGALS3BP in hepatocellular carcinoma (HCC) and its relationship with angiogenesis and the NOTCH signaling pathway. qRT-PCR, Western blot (WB), and ELISA were performed to validate the mRNA, protein expression, and secretion levels of LGALS3BP. The effects of LGALS3BP on HCC cell viability, proliferation, and invasion were assessed using CCK-8, colony formation, and Transwell assays. A co-culture system of HCC cells and human umbilical vein endothelial cells was established to evaluate pro-angiogenic effects. qRT-PCR and WB were used to detect mRNA and protein expression of the NOTCH pathway. Bioinformatics analysis combined with Co-IP and IF validated the interaction between UGGT1 and LGALS3BP. N-glycosylation sites of LGALS3BP were analyzed, and the role of UGGT1 in LGALS3BP N-glycosylation was investigated through protein stability assays, WB, and ELISA. The impact of UGGT1-mediated LGALS3BP N-glycosylation on the NOTCH pathway and HCC angiogenesis was explored via qRT-PCR, WB, functional cell assays, and co-culture experiments. Finally, in vivo validation was conducted using a xenograft tumor model. Results LGALS3BP was significantly overexpressed in HCC tissues and cells. Knockdown of LGALS3BP inhibited HCC cell proliferation and invasion, reduced VEGF-A expression, and suppressed tube formation in human umbilical-vein endothelial cells. Mechanistically, UGGT1 maintained LGALS3BP protein stability by mediating its N-glycosylation modification. LGALS3BP promoted VEGF-A expression by activating the NOTCH pathway. Animal experiments demonstrated that UGGT1 knockdown significantly inhibited tumor growth and angiogenesis, an effect that could be partially rescued by LGALS3BP overexpression. Conclusion This study reveals that UGGT1 promotes angiogenesis in HCC by mediating N-glycosylation of LGALS3BP and activating the NOTCH/VEGF-A signaling axis, providing a potential target for anti-angiogenic therapy in HCC. liver cancer UGGT1 LGALS3BP N-glycosylation modification angiogenesis Full Text Additional Declarations No competing interests reported. Table 1 to 3 are available in the Supplementary Files section. Supplementary Files SupplementaryFigure1.tif Supplementary Figure 1. Analysis of protein interaction between UGGT1 and LGALS3BP using the BioGRID website. Tables.docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 10 Feb, 2026 Reviews received at journal 10 Feb, 2026 Reviewers agreed at journal 19 Jan, 2026 Reviews received at journal 27 Dec, 2025 Reviewers agreed at journal 08 Dec, 2025 Reviewers agreed at journal 12 Nov, 2025 Reviewers invited by journal 12 Nov, 2025 Editor assigned by journal 07 Nov, 2025 Submission checks completed at journal 07 Nov, 2025 First submitted to journal 06 Nov, 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-8052281","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":549182771,"identity":"a171ebb3-b0a8-4e60-8c73-abb881a40a22","order_by":0,"name":"Benzhen He","email":"","orcid":"","institution":"the Affiliated Hospital of Shaoxing University","correspondingAuthor":false,"prefix":"","firstName":"Benzhen","middleName":"","lastName":"He","suffix":""},{"id":549182772,"identity":"356c2c9d-f20b-4c0a-b632-7e7c2f8dadfe","order_by":1,"name":"Liang Wang","email":"","orcid":"","institution":"the Affiliated Hospital of Shaoxing University","correspondingAuthor":false,"prefix":"","firstName":"Liang","middleName":"","lastName":"Wang","suffix":""},{"id":549182773,"identity":"cd43cdc3-a75c-48d1-97e2-c5b6bc9407d5","order_by":2,"name":"Jiadong Xia","email":"","orcid":"","institution":"the Affiliated Hospital of Shaoxing University","correspondingAuthor":false,"prefix":"","firstName":"Jiadong","middleName":"","lastName":"Xia","suffix":""},{"id":549182774,"identity":"52fb28c0-aa48-45ec-ae06-f446806b59c2","order_by":3,"name":"Xinggang Xu","email":"","orcid":"","institution":"the Affiliated Hospital of Shaoxing University","correspondingAuthor":false,"prefix":"","firstName":"Xinggang","middleName":"","lastName":"Xu","suffix":""},{"id":549182775,"identity":"b5419919-9a6d-4a97-a2e7-e76b2b280b10","order_by":4,"name":"Jianguo Xu","email":"","orcid":"","institution":"Zhejiang Provincial People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Jianguo","middleName":"","lastName":"Xu","suffix":""},{"id":549182776,"identity":"e6a94d82-bcdf-4716-9c90-564e2cb33dfb","order_by":5,"name":"Geqiong Xiao","email":"","orcid":"","institution":"the Affiliated Hospital of Shaoxing University","correspondingAuthor":false,"prefix":"","firstName":"Geqiong","middleName":"","lastName":"Xiao","suffix":""},{"id":549182777,"identity":"ca63c6d6-7f5a-45f3-86be-4352e8adc64b","order_by":6,"name":"Jiamin Sheng","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzElEQVRIiWNgGAWjYBACAwbGBgYGHgk5fmbmww9I0CJjYSzZzpZmQKQWELCpSDQ4z6MgQZQWc/bDzS8+5EgkGB/mAeqvsYkmqMWyJ7HNcsYZiTyzw7wHHjAcS8ttIOiwA4ltxrw9EsVmh/kSDBgbDhOh5fzDNuO//yQSNzfzGEgQp+VGYvNjYCAnbmAmVovljIdtjD08EsYSh4GBnECMX8z50x9/+MFTJ8fff/jwgw81NoS1AAEbIjoSiFAOAswfiFQ4CkbBKBgFIxUAAAoSPzvma/tEAAAAAElFTkSuQmCC","orcid":"","institution":"the Affiliated Hospital of Shaoxing University","correspondingAuthor":true,"prefix":"","firstName":"Jiamin","middleName":"","lastName":"Sheng","suffix":""}],"badges":[],"createdAt":"2025-11-07 02:38:22","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8052281/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8052281/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":96658215,"identity":"5e97eb99-e239-4102-b579-7f08e7760360","added_by":"auto","created_at":"2025-11-24 17:35:59","extension":"tif","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":9516196,"visible":true,"origin":"","legend":"","description":"","filename":"Figure1.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/c8599b496b1351d2f94d6e63.tif"},{"id":96658212,"identity":"cf315587-8a2d-44ad-9ee1-d216a8960f37","added_by":"auto","created_at":"2025-11-24 17:35:59","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":90856,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.docx","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/86ae36245edeef09d0f0dd7e.docx"},{"id":96658210,"identity":"dbf360b4-1478-42b2-be5e-b1cd65f5a947","added_by":"auto","created_at":"2025-11-24 17:35:59","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":15801,"visible":true,"origin":"","legend":"","description":"","filename":"Tables.docx","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/eacd2a3620ba83d083ea3cf0.docx"},{"id":96658217,"identity":"30654c6b-9cee-4b16-809f-6b08d845fd13","added_by":"auto","created_at":"2025-11-24 17:35:59","extension":"tif","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":10237408,"visible":true,"origin":"","legend":"","description":"","filename":"Figure2.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/dc384127665a750c48e38fd0.tif"},{"id":96658222,"identity":"012c4b36-43ce-4f4a-bcbf-a5274d842089","added_by":"auto","created_at":"2025-11-24 17:35:59","extension":"tif","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":2714104,"visible":true,"origin":"","legend":"","description":"","filename":"Figure3.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/ebe43b455ca62480d0089a1e.tif"},{"id":96658223,"identity":"e5878a8e-d43a-4b50-9135-48bd33c1eb46","added_by":"auto","created_at":"2025-11-24 17:35:59","extension":"tif","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":7800700,"visible":true,"origin":"","legend":"","description":"","filename":"Figure4.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/cd2ced01e30feb5f59ee9eb0.tif"},{"id":96709829,"identity":"5b509bc6-d71d-4b52-8b88-c74299c654f4","added_by":"auto","created_at":"2025-11-25 10:09:43","extension":"tif","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":11759840,"visible":true,"origin":"","legend":"","description":"","filename":"Figure5.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/9c39293c6d309d6155ffbeff.tif"},{"id":96658216,"identity":"34bdb4eb-ccc3-4038-821f-29f34a402c9a","added_by":"auto","created_at":"2025-11-24 17:35:59","extension":"json","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":9096,"visible":true,"origin":"","legend":"","description":"","filename":"c44ffcd757ed42c4881481adbc24af32.json","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/4c765f2561fda5d896b8aa45.json"},{"id":96658231,"identity":"1b93318d-97e9-44f4-b363-011d5969d794","added_by":"auto","created_at":"2025-11-24 17:36:00","extension":"tif","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":20569432,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryFigure1.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/f830f47f972955ffb981dd4e.tif"},{"id":96709855,"identity":"442dba3a-eaa7-44c8-854a-e08891a9d3e5","added_by":"auto","created_at":"2025-11-25 10:09:44","extension":"xml","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":116437,"visible":true,"origin":"","legend":"","description":"","filename":"c44ffcd757ed42c4881481adbc24af321enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/aaf2abaffd248d697b64a694.xml"},{"id":96658237,"identity":"27bcd01c-f1a0-4314-9fb6-8bf338333d6d","added_by":"auto","created_at":"2025-11-24 17:36:00","extension":"tif","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":9516196,"visible":true,"origin":"","legend":"","description":"","filename":"Figure1.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/e1756346d9acfbcc9c681031.tif"},{"id":96709807,"identity":"04478a0d-16f8-4a9c-ac13-9ef716b573a0","added_by":"auto","created_at":"2025-11-25 10:09:42","extension":"tif","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":10237408,"visible":true,"origin":"","legend":"","description":"","filename":"Figure2.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/80b479a65af1b3a6669a2955.tif"},{"id":96709945,"identity":"2c911681-4b0b-49d8-9acb-be3229437791","added_by":"auto","created_at":"2025-11-25 10:09:48","extension":"tif","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":2714104,"visible":true,"origin":"","legend":"","description":"","filename":"Figure3.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/b27bd53506595fd911157eae.tif"},{"id":96658230,"identity":"cee3e495-a779-4242-8154-5bae4818539f","added_by":"auto","created_at":"2025-11-24 17:36:00","extension":"tif","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":7800700,"visible":true,"origin":"","legend":"","description":"","filename":"Figure4.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/12f0c4a5bfe45a262f625849.tif"},{"id":96658233,"identity":"a49f40d0-1e6e-4c80-888c-3ae1870f89de","added_by":"auto","created_at":"2025-11-24 17:36:00","extension":"tif","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":11759840,"visible":true,"origin":"","legend":"","description":"","filename":"Figure5.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/657bba254e7709080630b719.tif"},{"id":96709872,"identity":"c9e4f257-04fc-4be1-abbd-5aeca8941546","added_by":"auto","created_at":"2025-11-25 10:09:44","extension":"png","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1227738,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure1.png","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/75631b3526e5aec691372c20.png"},{"id":96709670,"identity":"5cb84846-3aa0-474c-bcec-238a09940220","added_by":"auto","created_at":"2025-11-25 10:09:30","extension":"png","order_by":16,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1503875,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure2.png","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/f47fa9c61ae94b5e0a7ad488.png"},{"id":96658234,"identity":"846604db-a520-4fa5-87d9-7f3aceaaf5ed","added_by":"auto","created_at":"2025-11-24 17:36:00","extension":"png","order_by":17,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":393651,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure3.png","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/d4c9337db67f520129d670a0.png"},{"id":96658221,"identity":"30ded465-e60a-40c7-a1da-93c88c52481d","added_by":"auto","created_at":"2025-11-24 17:35:59","extension":"png","order_by":18,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1266143,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure4.png","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/e47897198916470df4563a92.png"},{"id":96658232,"identity":"afbfb440-1a49-4ae9-a398-bf39ad315a49","added_by":"auto","created_at":"2025-11-24 17:36:00","extension":"png","order_by":19,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1610259,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure5.png","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/7c7fd251b357b75095050c20.png"},{"id":96658218,"identity":"00a40c03-fbda-4512-ac3e-f86a69eaf9fb","added_by":"auto","created_at":"2025-11-24 17:35:59","extension":"xml","order_by":20,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":114699,"visible":true,"origin":"","legend":"","description":"","filename":"c44ffcd757ed42c4881481adbc24af321structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/aa72064938f5bf3c79b3150c.xml"},{"id":96709771,"identity":"afbe62a4-00a8-4bf0-90dd-cca41531cfb0","added_by":"auto","created_at":"2025-11-25 10:09:37","extension":"html","order_by":21,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":126023,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/3d1ec1718ec41a6debe8a392.html"},{"id":96712728,"identity":"47ffe4e7-cb73-49c3-962b-b2a3a29287f5","added_by":"auto","created_at":"2025-11-25 10:16:18","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1522486,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1_covered_463662ff-f69f-4daa-bba4-7dc112d84741.pdf"},{"id":96658235,"identity":"245da31f-88c8-4c8f-b376-2eae7e651b28","added_by":"auto","created_at":"2025-11-24 17:36:00","extension":"tif","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":20569432,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSupplementary Figure 1. \u003c/strong\u003eAnalysis of protein interaction between UGGT1 and LGALS3BP using the BioGRID website.\u003c/p\u003e","description":"","filename":"SupplementaryFigure1.tif","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/bf88ed27dca82bee1782ddca.tif"},{"id":96658214,"identity":"95ba13a6-1bcc-49a1-a9cd-61d5aebad8f5","added_by":"auto","created_at":"2025-11-24 17:35:59","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":15801,"visible":true,"origin":"","legend":"","description":"","filename":"Tables.docx","url":"https://assets-eu.researchsquare.com/files/rs-8052281/v1/353e3e9b79c5c39b40d9b3d6.docx"}],"financialInterests":"\u003cp\u003eNo competing interests reported.\u003c/p\u003e\n\u003cp\u003eTable 1 to 3 are available in the Supplementary Files section.\u003c/p\u003e","formattedTitle":"UGGT1 mediates N-glycosylation of LGALS3BP to induce angiogenesis in liver cancer via the NOTCH signaling pathway","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"clinical-proteomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"clip","sideBox":"Learn more about [Clinical Proteomics](http://clinicalproteomicsjournal.biomedcentral.com/)","snPcode":"12014","submissionUrl":"https://submission.nature.com/new-submission/12014/3","title":"Clinical Proteomics","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"liver cancer, UGGT1, LGALS3BP, N-glycosylation modification, angiogenesis","lastPublishedDoi":"10.21203/rs.3.rs-8052281/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8052281/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003eLiver cancer (LC) is one of the leading causes of cancer-related deaths worldwide. Its complex vascular regulatory network and drug resistance severely limit clinical efficacy. Although LGALS3BP is highly expressed in various malignant tumors, its biological function and molecular regulatory mechanisms in LC angiogenesis remain to be elucidated.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eBioinformatics analysis was used to examine the expression of LGALS3BP in hepatocellular carcinoma (HCC) and its relationship with angiogenesis and the NOTCH signaling pathway. qRT-PCR, Western blot (WB), and ELISA were performed to validate the mRNA, protein expression, and secretion levels of LGALS3BP. The effects of LGALS3BP on HCC cell viability, proliferation, and invasion were assessed using CCK-8, colony formation, and Transwell assays. A co-culture system of HCC cells and human umbilical vein endothelial cells was established to evaluate pro-angiogenic effects. qRT-PCR and WB were used to detect mRNA and protein expression of the NOTCH pathway. Bioinformatics analysis combined with Co-IP and IF validated the interaction between UGGT1 and LGALS3BP. N-glycosylation sites of LGALS3BP were analyzed, and the role of UGGT1 in LGALS3BP N-glycosylation was investigated through protein stability assays, WB, and ELISA. The impact of UGGT1-mediated LGALS3BP N-glycosylation on the NOTCH pathway and HCC angiogenesis was explored via qRT-PCR, WB, functional cell assays, and co-culture experiments. Finally, \u003cem\u003ein vivo\u003c/em\u003e validation was conducted using a xenograft tumor model.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eLGALS3BP was significantly overexpressed in HCC tissues and cells. Knockdown of LGALS3BP inhibited HCC cell proliferation and invasion, reduced VEGF-A expression, and suppressed tube formation in human umbilical-vein endothelial cells. Mechanistically, UGGT1 maintained LGALS3BP protein stability by mediating its N-glycosylation modification. LGALS3BP promoted VEGF-A expression by activating the NOTCH pathway. Animal experiments demonstrated that UGGT1 knockdown significantly inhibited tumor growth and angiogenesis, an effect that could be partially rescued by LGALS3BP overexpression.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eThis study reveals that UGGT1 promotes angiogenesis in HCC by mediating N-glycosylation of LGALS3BP and activating the NOTCH/VEGF-A signaling axis, providing a potential target for anti-angiogenic therapy in HCC.\u003c/p\u003e","manuscriptTitle":"UGGT1 mediates N-glycosylation of LGALS3BP to induce angiogenesis in liver cancer via the NOTCH signaling pathway","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-24 17:35:54","doi":"10.21203/rs.3.rs-8052281/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-10T12:27:36+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-10T12:15:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"338510492963108421593477286286640183577","date":"2026-01-19T16:08:01+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-27T06:45:34+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"194916628584371141154516566199326207386","date":"2025-12-08T09:56:14+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"151235999912557873020154165225783750892","date":"2025-11-12T15:29:58+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-12T15:05:46+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-07T08:38:25+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-07T08:37:04+00:00","index":"","fulltext":""},{"type":"submitted","content":"Clinical Proteomics","date":"2025-11-07T02:31:09+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"clinical-proteomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"clip","sideBox":"Learn more about [Clinical Proteomics](http://clinicalproteomicsjournal.biomedcentral.com/)","snPcode":"12014","submissionUrl":"https://submission.nature.com/new-submission/12014/3","title":"Clinical Proteomics","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"d3267445-fced-4d7c-bbd8-1eb6ff7fe047","owner":[],"postedDate":"November 24th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-04-02T08:09:34+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-24 17:35:54","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8052281","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8052281","identity":"rs-8052281","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}