Comprehensive bioinformatics analysis reveals CLCA1 and ZG16 as predictive biomarkers of malignant progression in colorectal cancer

Comprehensive bioinformatics analysis reveals CLCA1 and ZG16 as predictive biomarkers of malignant progression in colorectal cancer | 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 Comprehensive bioinformatics analysis reveals CLCA1 and ZG16 as predictive biomarkers of malignant progression in colorectal cancer Jialin Zhang, Xinyu Wang, Ziqiang Wang, Xiaona Hao, Yuyun Li, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4930170/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Colorectal cancer (CRC) is one of the most common malignant tumors. CLCA1 and ZG16 are lowly expressed in CRC, and we wanted to investigate whether they could be prognostic biomarkers for the malignant progression of CRC. Methods 12,195 DEGs and 12,071 DEGs were identified through the GSE39582 dataset and TCGA dataset, and then 50 coexisting genes were selected for further analysis using Venn diagrams. These 50 DEGs were then subjected to GO and KEGG functional enrichment analyses, along with genome-wide GSEA. the first 5 core genes were identified and visualized using Cytoscape through the PPI network. Then the expression of ZG16 and CLCA1 in normal and tumor tissues were analyzed using GSE39582 and TCGA datasets, and correlation analysis, and survival analysis were performed. The expression of ZG16 and CLCA1 in CRC cells was verified by qRT-PCR, and cell proliferation, migration, and invasion abilities were detected by CCK-8, scratch assay, clone formation assay, and Transwell assay. Results The expression levels of ZG16 and CLCA1 were significantly lower in tissues from CRC patients than in normal tissues. Survival analysis showed that low expression of ZG16 and CLCA1 was associated with poor survival outcomes. Multifactorial analysis showed that low expression of ZG16 and CLCA1 was an independent risk factor affecting tumor prognosis. Cellular experiments showed that cell proliferation, migration, and invasion were inhibited after overexpression of ZG16 and CLCA1. Correlation analysis showed that ZG16 and CLCA1 expression levels were positively correlated and the correlation was statistically significant. GSEA enrichment analysis based on CLCA1-related genes and ZG16-related genes (FDR < 0.25, P < 0.05) revealed that the related genes of both genes were closely related to the GNRH SINALINGPATHWAYES pathway. Conclusion CLCA1 and ZG16, which are lowly expressed in CRC tissues, are associated with poor prognosis of CRC and may be one of the markers for diagnostic screening and prediction of prognostic outcome in CRC. Meanwhile, CLCA1 and ZG16 may also be new targets for tumor immunotherapy. CLCA1 ZG16 colorectal cancer biomarker gene set enrichment analysis prognosis Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted 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. 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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-4930170","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":351118683,"identity":"73b6ac4d-7339-4005-8d19-eeb3e9d02e2c","order_by":0,"name":"Jialin Zhang","email":"","orcid":"","institution":"Bengbu Medical University, Bengbu Medical University","correspondingAuthor":false,"prefix":"","firstName":"Jialin","middleName":"","lastName":"Zhang","suffix":""},{"id":351118684,"identity":"2628b0b0-6787-4940-ac6c-21d491464df1","order_by":1,"name":"Xinyu Wang","email":"","orcid":"","institution":"Bengbu Medical University, Bengbu Medical University","correspondingAuthor":false,"prefix":"","firstName":"Xinyu","middleName":"","lastName":"Wang","suffix":""},{"id":351118685,"identity":"6361f29b-da1f-4512-9221-3a2c37dd6175","order_by":2,"name":"Ziqiang Wang","email":"","orcid":"","institution":"Bengbu Medical University, Bengbu Medical University","correspondingAuthor":false,"prefix":"","firstName":"Ziqiang","middleName":"","lastName":"Wang","suffix":""},{"id":351118686,"identity":"b2996ee7-5a6c-4d09-8f08-cc47f7c9512e","order_by":3,"name":"Xiaona Hao","email":"","orcid":"","institution":"Bengbu Medical University, Bengbu Medical University","correspondingAuthor":false,"prefix":"","firstName":"Xiaona","middleName":"","lastName":"Hao","suffix":""},{"id":351118687,"identity":"34acc3be-2aa1-416a-bcf9-825fa44979b9","order_by":4,"name":"Yuyun Li","email":"","orcid":"","institution":"Bengbu Medical University, Bengbu Medical University","correspondingAuthor":false,"prefix":"","firstName":"Yuyun","middleName":"","lastName":"Li","suffix":""},{"id":351118688,"identity":"82c84b30-6316-49d2-8264-3d3975617bae","order_by":5,"name":"Yingjie Zhang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAtUlEQVRIiWNgGAWjYBACfvnDBx9+qLCRI16L5Ay2ZGOJM2nGxGsxuMFjJsHbdiixgXhbZvcYSEicOZDedzyB8cPHHCK08MscKzAoqLiTO/PMA2bJmduIsaUheUOCxJlnuRtuJLAx8xKjxeBAgsEB3rbD6QbEa7mRYtgA1JJAvBbJnmPJzMBANpx55mEzcX7hZ28+/hMYlfJ8x5MPfvhIjBYEOEBC1MC0JJCqYxSMglEwCkYKAAD6Oj/ZO4oY/QAAAABJRU5ErkJggg==","orcid":"","institution":"Bengbu Medical University, Bengbu Medical University","correspondingAuthor":true,"prefix":"","firstName":"Yingjie","middleName":"","lastName":"Zhang","suffix":""}],"badges":[],"createdAt":"2024-08-17 14:14:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4930170/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4930170/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":65238028,"identity":"be76ba41-9f0a-474d-9267-9d8c667a77d9","added_by":"auto","created_at":"2024-09-25 06:09:09","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1460816,"visible":true,"origin":"","legend":"","description":"","filename":"Article.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4930170/v1_covered_330ec1d3-cb9a-41cb-8d62-f67d103236a1.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Comprehensive bioinformatics analysis reveals CLCA1 and ZG16 as predictive biomarkers of malignant progression in colorectal cancer","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"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":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"CLCA1, ZG16, colorectal cancer, biomarker, gene set enrichment analysis, prognosis","lastPublishedDoi":"10.21203/rs.3.rs-4930170/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4930170/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eColorectal cancer (CRC) is one of the most common malignant tumors. CLCA1 and ZG16 are lowly expressed in CRC, and we wanted to investigate whether they could be prognostic biomarkers for the malignant progression of CRC.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003e12,195 DEGs and 12,071 DEGs were identified through the GSE39582 dataset and TCGA dataset, and then 50 coexisting genes were selected for further analysis using Venn diagrams. These 50 DEGs were then subjected to GO and KEGG functional enrichment analyses, along with genome-wide GSEA. the first 5 core genes were identified and visualized using Cytoscape through the PPI network. Then the expression of ZG16 and CLCA1 in normal and tumor tissues were analyzed using GSE39582 and TCGA datasets, and correlation analysis, and survival analysis were performed. The expression of ZG16 and CLCA1 in CRC cells was verified by qRT-PCR, and cell proliferation, migration, and invasion abilities were detected by CCK-8, scratch assay, clone formation assay, and Transwell assay.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThe expression levels of ZG16 and CLCA1 were significantly lower in tissues from CRC patients than in normal tissues. Survival analysis showed that low expression of ZG16 and CLCA1 was associated with poor survival outcomes. Multifactorial analysis showed that low expression of ZG16 and CLCA1 was an independent risk factor affecting tumor prognosis. Cellular experiments showed that cell proliferation, migration, and invasion were inhibited after overexpression of ZG16 and CLCA1. Correlation analysis showed that ZG16 and CLCA1 expression levels were positively correlated and the correlation was statistically significant. GSEA enrichment analysis based on CLCA1-related genes and ZG16-related genes (FDR\u0026thinsp;\u0026lt;\u0026thinsp;0.25, P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) revealed that the related genes of both genes were closely related to the GNRH SINALINGPATHWAYES pathway.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eCLCA1 and ZG16, which are lowly expressed in CRC tissues, are associated with poor prognosis of CRC and may be one of the markers for diagnostic screening and prediction of prognostic outcome in CRC. Meanwhile, CLCA1 and ZG16 may also be new targets for tumor immunotherapy.\u003c/p\u003e","manuscriptTitle":"Comprehensive bioinformatics analysis reveals CLCA1 and ZG16 as predictive biomarkers of malignant progression in colorectal cancer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-09-23 17:00:58","doi":"10.21203/rs.3.rs-4930170/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b3a80834-a2db-4474-9204-b0d3422196f6","owner":[],"postedDate":"September 23rd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-09-25T06:08:33+00:00","versionOfRecord":[],"versionCreatedAt":"2024-09-23 17:00:58","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4930170","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4930170","identity":"rs-4930170","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}