{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T11:04:10Z","timestamp":1785409450837,"version":"3.56.0"},"reference-count":58,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T00:00:00Z","timestamp":1780272000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Outstanding Youth Research Project of Anhui, Department of Education","award":["2022AH020042"],"award-info":[{"award-number":["2022AH020042"]}]},{"name":"Major Scientific Research Project of Universities in Anhui province","award":["2024AH040143"],"award-info":[{"award-number":["2024AH040143"]}]},{"name":"Major Scientific Research Project of Universities in Anhui province","award":["2024AH040146"],"award-info":[{"award-number":["2024AH040146"]}]},{"name":"key projects of Anhui Provincial Department of Education","award":["2023sdxx027"],"award-info":[{"award-number":["2023sdxx027"]}]},{"name":"key projects of Anhui Provincial Department of Education","award":["2024AH052728"],"award-info":[{"award-number":["2024AH052728"]}]},{"award":["2023sdxx027"],"award-info":[{"award-number":["2023sdxx027"]}],"id":[{"id":"https:\/\/ror.org\/012m7k033","id-type":"ROR","asserted-by":"publisher"}]},{"award":["2024AH052728"],"award-info":[{"award-number":["2024AH052728"]}],"id":[{"id":"https:\/\/ror.org\/012m7k033","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Training Action Project of Anhui Provincial Education Department of Young and Middle-aged Teachers","award":["JNFX2023020"],"award-info":[{"award-number":["JNFX2023020"]}]},{"name":"Training Action Project of Anhui Provincial Education Department of Young and Middle-aged Teachers","award":["JNFX2024023"],"award-info":[{"award-number":["JNFX2024023"]}]},{"name":"Training Action Project of Anhui Provincial Education Department of Young and Middle-aged Teachers","award":["2023sdxx027"],"award-info":[{"award-number":["2023sdxx027"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Objective: To elucidate the role of cytoskeleton-associated protein 2 (CKAP2) in gastric cancer (GC) progression and evaluate its prognostic and potential protective significance. Methods: The candidate genes for GC were identified using differential expression and weighted gene co-expression network analysis (WGCNA). A panel of machine learning algorithms was applied for obtaining the key genes and yielding the hub target gene CKAP2 through protein\u2013protein interaction (PPI) networks and single-cell RNA sequencing analysis. Further, the functional role of CKAP2 was explored through single-cell virtual knockout and pathway enrichment analysis. Meanwhile, the survival analysis and Mendelian randomization (MR) were used to evaluate the clinical relevance and causality of CKAP2. Finally, the expression of CKAP2 was further validated in clinical tissues and GC cell lines through Western blot. Results: A total of 20 candidate genes were identified, of which 8 were identified as hub genes through machine learning. Integrative PPI and single-cell analyses ultimately identified CKAP2 as the key gene. Virtual knockout analysis showed that the related differentially expressed genes were significantly enriched in the 5-HT pathway. Survival analysis demonstrated that elevated CKAP2 expression was associated with improved prognosis. MR analysis further suggested that CKAP2 might act as a protective factor, while 5-HT was associated with an increased risk of GC. Experimental validation confirmed that CKAP2 is significantly upregulated in GC tissues and cell lines. Conclusions: CKAP2 is a potential prognostic biomarker and protective factor in GC, possibly exerting its effects through regulation of the 5-HT pathway. These findings provide new insights into the mechanisms and potential therapeutic targets of GC.<\/jats:p>","DOI":"10.3390\/molecules31111901","type":"journal-article","created":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T15:07:52Z","timestamp":1780326472000},"page":"1901","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Integrated Single-Cell Virtual Knockout and Machine Learning Analyses Reveal a Protective Role of CKAP2 in Gastric Cancer"],"prefix":"10.3390","volume":"31","author":[{"given":"Jianhua","family":"Yang","sequence":"first","affiliation":[{"name":"School of Medical Informatics Engineering, Anhui University of Chinese Medicine, Hefei 230012, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-5552-4083","authenticated-orcid":false,"given":"Zheng","family":"Qiu","sequence":"additional","affiliation":[{"name":"School of Medical Informatics Engineering, Anhui University of Chinese Medicine, Hefei 230012, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenchao","family":"Song","sequence":"additional","affiliation":[{"name":"School of Medical Informatics Engineering, Anhui University of Chinese Medicine, Hefei 230012, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xing","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Medical Informatics Engineering, Anhui University of Chinese Medicine, Hefei 230012, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting","family":"Ouyang","sequence":"additional","affiliation":[{"name":"School of Medical Informatics Engineering, Anhui University of Chinese Medicine, Hefei 230012, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianhua","family":"Shu","sequence":"additional","affiliation":[{"name":"School of Medical Informatics Engineering, Anhui University of Chinese Medicine, Hefei 230012, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8462-3322","authenticated-orcid":false,"given":"Jinghui","family":"Wang","sequence":"additional","affiliation":[{"name":"Anhui Provincial Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine, Hefei 230012, China"},{"name":"School of Integrated Chinese and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinfeng","family":"Yang","sequence":"additional","affiliation":[{"name":"School of Medical Informatics Engineering, Anhui University of Chinese Medicine, Hefei 230012, China"},{"name":"Anhui Provincial Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine, Hefei 230012, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2661","DOI":"10.1002\/ptr.7866","article-title":"Natural products targeting signaling pathways associated with regulated cell death in gastric cancer: Recent advances and perspectives","volume":"37","author":"Hu","year":"2023","journal-title":"Phytother. 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