Salvianolic acid B inhibits the growth and metastasis of A549 lung cancer cells through the NDRG2/PTEN pathway by inducing oxidative stress

Salvianolic acid B inhibits the growth and metastasis of A549 lung cancer cells through the NDRG2/PTEN pathway by inducing oxidative stress | 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 Salvianolic acid B inhibits the growth and metastasis of A549 lung cancer cells through the NDRG2/PTEN pathway by inducing oxidative stress Ye Yang, Lei Huang, Jie Gao, Bingjun Qian This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4276264/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 07 Jun, 2024 Read the published version in Medical Oncology → Version 1 posted 7 You are reading this latest preprint version Abstract Salvianolic acid B (Sal B) has demonstrated anticancer activity against various types of cancer. However, the underlying mechanism of Sal B-mediated anticancer effects remains incompletely understood. This study aims to investigate the impact of Sal B on the growth and metastasis of human A549 lung cells, as well as elucidate its potential mechanisms. In this study, different concentrations of Sal B were administered to A549 cells. The effects on migration and invasion abilities were assessed using MTT, wound healing, and transwell assays. Flow cytometry analysis was employed to evaluate Sal B-induced apoptosis in A549 cells. Western blotting and immunohistochemistry were conducted to measure the expression levels of cleaved caspase 3 (an apoptosis marker) and E-cadherin (a metastasis marker). Commercial kits were utilized for detecting intracellular reactive oxygen species (ROS) and NAD + . Additionally, a xenograft model with transplanted A549 tumors was employed to assess the antitumor effect of Sal B in vivo. The expression levels of NDRG2, p-PTEN, and p-AKT were determined through western blotting. Our findings demonstrate that Sal B effectively inhibits proliferation, migration, and invasion in A549 cells while inducing dose-dependent apoptosis. These apoptotic responses and inhibition of tumor cell metastasis are accompanied by alterations in intracellular ROS levels and NAD + /NADH ratio. Furthermore, our in vivo experiment reveals that Sal B significantly suppresses A549 tumor growth compared to an untreated control group while promoting increased cleavage of caspase-3. Importantly, we observe that Sal B upregulates NDRG2 expression while downregulating p-PTEN and p-AKT expressions. Collectively, our results provide compelling evidence supporting the ability of Sal B to inhibit both growth and metastasis in A549 lung cancer cells through oxidative stress modulation as well as involvement of the NDRG2/PTEN/AKT pathway. Salvianolic acid B A549 NDRG2 p-PTEN growth metastasis ROS Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 07 Jun, 2024 Read the published version in Medical Oncology → Version 1 posted Editorial decision: Revision requested 12 May, 2024 Reviews received at journal 10 May, 2024 Reviewers agreed at journal 01 May, 2024 Reviewers invited by journal 29 Apr, 2024 Submission checks completed at journal 17 Apr, 2024 Editor assigned by journal 17 Apr, 2024 First submitted to journal 16 Apr, 2024 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. 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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-4276264","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":293897393,"identity":"1d5a1cdf-8117-4e04-8083-4e3f78e9e9b0","order_by":0,"name":"Ye Yang","email":"","orcid":"","institution":"Jiangsu Vocational College of Medicine","correspondingAuthor":false,"prefix":"","firstName":"Ye","middleName":"","lastName":"Yang","suffix":""},{"id":293897394,"identity":"93c1a7a1-13f1-4790-876c-bcabee8b4c98","order_by":1,"name":"Lei Huang","email":"","orcid":"","institution":"Jiangsu Vocational College of Medicine","correspondingAuthor":false,"prefix":"","firstName":"Lei","middleName":"","lastName":"Huang","suffix":""},{"id":293897395,"identity":"db8c5a54-cce1-477d-870d-8c5e7bb68de2","order_by":2,"name":"Jie Gao","email":"","orcid":"","institution":"Jiangsu Provincial Hospital of Traditional Chinese Medicine","correspondingAuthor":false,"prefix":"","firstName":"Jie","middleName":"","lastName":"Gao","suffix":""},{"id":293897396,"identity":"dca32457-9412-40a8-880d-c39f19bd880e","order_by":3,"name":"Bingjun Qian","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA30lEQVRIie3RvQrCMBDA8QtCJ8W1S3yGg0CLIL5LEZwdHQNCJz9WfYtIQRyvBOzSB1BcXOzm4KagaFRc046C+WdIhvwgRwBcrh8MgcyCFpu9NhOTFYlgC1mdvBNMfUQFEjK9pOsaeZJmR3WBDldUKw420pabQTrJUawoD3Zj6AtFXojWh1GO1IhvkSHeHkBHiuqeX0bSe4xRIrPCkEc1ohuGKKDAEConr1k0j1HM37NgTyy0F1hJ6OvkfIqRz+ZZsb0Mu3yajQorAZ/we4DPN9Ws901Nid9D2VWXy+X6156lC1rJTywRjAAAAABJRU5ErkJggg==","orcid":"","institution":"Jiangsu Vocational College of Medicine","correspondingAuthor":true,"prefix":"","firstName":"Bingjun","middleName":"","lastName":"Qian","suffix":""}],"badges":[],"createdAt":"2024-04-16 13:13:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4276264/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4276264/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s12032-024-02413-6","type":"published","date":"2024-06-07T14:47:43+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":58822031,"identity":"106eee9e-cbdc-4bac-97ff-37e243aebf40","added_by":"auto","created_at":"2024-06-21 16:25:55","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":869099,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4276264/v1_covered_fecbe47a-22ea-46d4-a7f6-927a351bee4e.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Salvianolic acid B inhibits the growth and metastasis of A549 lung cancer cells through the NDRG2/PTEN pathway by inducing oxidative stress","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":"medical-oncology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"medo","sideBox":"Learn more about [Medical Oncology](https://www.springer.com/journal/12032)","snPcode":"12032","submissionUrl":"https://submission.nature.com/new-submission/12032/3","title":"Medical Oncology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Salvianolic acid B, A549, NDRG2, p-PTEN, growth, metastasis, ROS","lastPublishedDoi":"10.21203/rs.3.rs-4276264/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4276264/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSalvianolic acid B (Sal B) has demonstrated anticancer activity against various types of cancer. However, the underlying mechanism of Sal B-mediated anticancer effects remains incompletely understood. This study aims to investigate the impact of Sal B on the growth and metastasis of human A549 lung cells, as well as elucidate its potential mechanisms. In this study, different concentrations of Sal B were administered to A549 cells. The effects on migration and invasion abilities were assessed using MTT, wound healing, and transwell assays. Flow cytometry analysis was employed to evaluate Sal B-induced apoptosis in A549 cells. Western blotting and immunohistochemistry were conducted to measure the expression levels of cleaved caspase 3 (an apoptosis marker) and E-cadherin (a metastasis marker). Commercial kits were utilized for detecting intracellular reactive oxygen species (ROS) and NAD\u003csup\u003e+\u003c/sup\u003e. Additionally, a xenograft model with transplanted A549 tumors was employed to assess the antitumor effect of Sal B in vivo. The expression levels of NDRG2, p-PTEN, and p-AKT were determined through western blotting. Our findings demonstrate that Sal B effectively inhibits proliferation, migration, and invasion in A549 cells while inducing dose-dependent apoptosis. These apoptotic responses and inhibition of tumor cell metastasis are accompanied by alterations in intracellular ROS levels and NAD\u003csup\u003e+\u003c/sup\u003e/NADH ratio. Furthermore, our in vivo experiment reveals that Sal B significantly suppresses A549 tumor growth compared to an untreated control group while promoting increased cleavage of caspase-3. Importantly, we observe that Sal B upregulates NDRG2 expression while downregulating p-PTEN and p-AKT expressions. 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