{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T09:02:29Z","timestamp":1789635749362,"version":"4.0.0"},"reference-count":29,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T00:00:00Z","timestamp":1789603200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background. Pancreatic ductal adenocarcinoma remains difficult to treat and has an exceptionally poor prognosis. While prior studies have noted antitumour properties for midazolam in pancreatic cancer, whether this response involves developmental signaling pathways is unclear. Here, we evaluated midazolam-treated MIA PaCa-2 cells to identify associated Wnt and Hedgehog transcriptional shifts and their systems-level context. Methods. MIA PaCa-2 cells were exposed to midazolam (20\u2013100 \u00b5M) for 24 h. We measured metabolic viability by MTT and apoptosis by Annexin V-FITC\/7-AAD flow cytometry at 60 \u00b5M. Expression of 18 targeted genes was quantified by RT-qPCR using ACTB for normalization. Changed genes were examined through STRING\/KEGG networks, TCGA-PAAD survival analysis, DGIdb annotations, and preliminary FZD7 molecular docking. Results. Midazolam reduced MTT-derived metabolic viability in a concentration-dependent manner (IC50 = 54.6 \u00b5M) and significantly increased Annexin V-positive apoptotic populations at 60 \u00b5M across three independent experiments. Eight transcripts changed significantly: CTNNB1, TGFBR2, WNT2, WNT7B, HHIP, and DHH declined, while CSNK1A1 and AXIN1 increased (all adjusted p &lt; 0.05). Network analysis highlighted CTNNB1 as the main hub, and high CTNNB1 expression correlated with shorter survival in TCGA-PAAD patients (p = 0.043; HR = 1.58, 95% CI: 1.01\u20132.47). Conclusions. Midazolam reduced metabolic viability and increased apoptosis in MIA PaCa-2 cells alongside coordinated expression changes in key Wnt and Hedgehog components. Because the required doses far exceed clinical sedation levels and pathway activity was not directly confirmed at the protein level, these findings should be regarded as a preliminary hypothesis rather than evidence of therapeutic efficacy.<\/jats:p>","DOI":"10.3390\/ph19091472","type":"journal-article","created":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T08:03:17Z","timestamp":1789632197000},"page":"1472","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Midazolam Reduces Metabolic Viability and Induces Apoptosis in MIA PaCa-2 Pancreatic Cancer Cells with Selective Transcriptional Modulation of Wnt and Hedgehog Components: A Preliminary In Vitro Study"],"prefix":"10.3390","volume":"19","author":[{"given":"Irmak Fato\u015f","family":"Korkmaz","sequence":"first","affiliation":[{"name":"Department of Anaesthesia, University of Health Science, Bah\u00e7elievler State Hospital, Istanbul 34668, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1311-6892","authenticated-orcid":false,"given":"Tugba","family":"Elgun","sequence":"additional","affiliation":[{"name":"Medical Biology, Faculty of Medicine, Biruni University, Istanbul 34015, T\u00fcrkiye"},{"name":"Biruni Advanced Technology Application and Research Center (B@MER), Biruni University, Istanbul 34015, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-1448-2739","authenticated-orcid":false,"given":"Ci\u011fdem","family":"Aktas","sequence":"additional","affiliation":[{"name":"Department of Genetics, Aziz Sancar Institute, Istanbul University, Istanbul 34452, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sajjad","family":"Eslamkhah","sequence":"additional","affiliation":[{"name":"Biruni Advanced Technology Application and Research Center (B@MER), Biruni University, Istanbul 34015, T\u00fcrkiye"},{"name":"Program of Pathology Laboratory Techniques, Vocational School, Biruni University, Istanbul 34015, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sevgi Kocyigit","family":"Sevinc","sequence":"additional","affiliation":[{"name":"Department of Biophysics, Faculty of Medicine, Kutahya Health Sciences University, Kutahya 43100, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6424-7411","authenticated-orcid":false,"given":"Asiye Gok","family":"Yurttas","sequence":"additional","affiliation":[{"name":"Department of Medical Biochemistry, Faculty of Medicine, Istanbul Atlas University, Istanbul 34403, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,17]]},"reference":[{"key":"ref_1","first-page":"450","article-title":"PLK1 downregulation prevents proliferation via the Wnt\/\u03b2-catenin pathway in cancer","volume":"17","author":"Ding","year":"2021","journal-title":"Arch. 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