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Therefore, there is an urgent need to develop new drugs to treat or prevent the prominent pathological subtype of esophageal cancer, esophageal squamous cell carcinoma (ESCC). Based upon the screening of drugs approved by the Food and Drug Administration, we discovered that Arbidol could effectively inhibit the proliferation of human ESCC in vitro. Next, we conducted a series of cell-based assays and found that Arbidol treatment inhibited the proliferation and colony formation ability of ESCC cells and promoted G1-phase cell cycle arrest. Phosphoproteomics experiments, in vitro kinase assays and pull-down assays were subsequently performed in order to identify the underlying growth inhibitory mechanism. We verified that Arbidol is a potential ataxia telangiectasia and Rad3-related (ATR) inhibitor via binding to ATR kinase to reduce the phosphorylation and activation of minichromosome maintenance protein 2 at Ser108. Finally, we demonstrated Arbidol had the inhibitory effect of ESCC in vivo by a patient-derived xenograft model. All together, Arbidol inhibits the proliferation of ESCC in vitro and in vivo through the DNA replication pathway and is associated with the cell cycle.<\/jats:p>","DOI":"10.7554\/elife.73953","type":"journal-article","created":{"date-parts":[[2022,9,9]],"date-time":"2022-09-09T06:56:12Z","timestamp":1662706572000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.73953","source":"Crossref","is-referenced-by-count":4,"title":["Arbidol inhibits human esophageal squamous cell carcinoma growth in vitro and in vivo through suppressing ataxia telangiectasia and Rad3-related protein kinase"],"prefix":"10.7554","volume":"11","author":[{"given":"Ning","family":"Yang","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04ypx8c21","id-type":"ROR","asserted-by":"publisher"}],"name":"Pathophysiology Department, The School of Basic Medical Sciences, Zhengzhou University","place":["Zhengzhou, 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