{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T23:33:19Z","timestamp":1783985599545,"version":"3.55.0"},"reference-count":33,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,8,23]],"date-time":"2025-08-23T00:00:00Z","timestamp":1755907200000},"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\/Objectives: Aristolochia ringens, a medicinal plant widely used in traditional medicine, has shown potential therapeutic applications. This study aimed to investigate the anticancer mechanism of action of its crude extract against human colorectal adenocarcinoma cells (Caco-2 and HT-29). Methods: Cell viability was assessed using the MTT assay to determine IC50 values. Immunofluorescence microscopy was used to examine nuclear morphology and microtubule integrity. Flow cytometry with PI staining was used for cell cycle analysis and Annexin V-FITC\/PI staining for apoptosis detection. Mitochondrial membrane potential was evaluated using JC-1 dye. Bioactivity-guided fractionation was performed via HPLC, and GC\u2013MS was used to profile active constituents. Results: The extract exhibited dose-dependent cytotoxicity with IC50 values below 30 \u00b5g\/mL in colon adenocarcinoma cell lines. Treated Caco-2 cells showed nuclear shrinkage and disrupted microtubules. PI-based flow cytometry revealed G1 phase arrest, and Annexin V-FITC\/PI staining indicated enhanced late apoptosis. JC-1 staining demonstrated mitochondrial depolarization. HPLC fractionation identified fractions 2 and 3 as active, and preliminary GC\u2013MS analysis tentatively annotated the presence of alkaloids, sesquiterpenes\/diterpenes, and steroidal compounds. Conclusions: A. ringens exerts anticancer effects through a mitochondria-mediated apoptotic pathway, involving G1 checkpoint arrest and cytoskeletal disruption. These findings provide the first integrated cellular and mechanistic evidence of its anticancer potential in colorectal cancer, supporting its promise as a source of novel therapeutic lead compounds.<\/jats:p>","DOI":"10.3390\/ph18091250","type":"journal-article","created":{"date-parts":[[2025,8,25]],"date-time":"2025-08-25T00:09:32Z","timestamp":1756080572000},"page":"1250","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Bioactivity-Guided Fractionation and Mechanistic Insights into Aristolochia ringens Root Extract-Induced G1 Phase Arrest and Mitochondria-Mediated Apoptosis in Human Colon Adenocarcinoma Cells"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5426-2581","authenticated-orcid":false,"given":"Saheed O.","family":"Anifowose","sequence":"first","affiliation":[{"name":"Zoology Department, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7362-2578","authenticated-orcid":false,"given":"Abdalrhaman M.","family":"Salih","sequence":"additional","affiliation":[{"name":"Botany and Microbiology Department, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-4949-0489","authenticated-orcid":false,"given":"Musa K.","family":"Oladejo","sequence":"additional","affiliation":[{"name":"Zoology Department, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmad","family":"Rady","sequence":"additional","affiliation":[{"name":"Zoology Department, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mobarak S.","family":"Al Mosallam","sequence":"additional","affiliation":[{"name":"Healthy Aging Research Institute, Health Sector, King Abdulaziz City for Science and Technology, Riyadh 11442, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hasan A.","family":"Aljohi","sequence":"additional","affiliation":[{"name":"Applied Genetics Technology Institute, King Abdulaziz City for Science and Technology, Riyadh 11442, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mansour I.","family":"Almansour","sequence":"additional","affiliation":[{"name":"Zoology Department, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saad Hussin","family":"Alkahtani","sequence":"additional","affiliation":[{"name":"Zoology Department, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8479-7875","authenticated-orcid":false,"given":"Ibrahim O.","family":"Alanazi","sequence":"additional","affiliation":[{"name":"Healthy Aging Research Institute, Health Sector, King Abdulaziz City for Science and Technology, Riyadh 11442, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Badr A.","family":"Al-Dahmash","sequence":"additional","affiliation":[{"name":"Zoology Department, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1038\/s41575-019-0189-8","article-title":"Global burden of colorectal cancer: Emerging trends, risk factors and prevention strategies","volume":"16","author":"Keum","year":"2019","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_2","first-page":"141","article-title":"Drug resistance and combating drug resistance in cancer","volume":"2","author":"Wang","year":"2019","journal-title":"Cancer Drug Resist."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"495","DOI":"10.4240\/wjgs.v15.i4.495","article-title":"Current and emerging therapeutic approaches for colorectal cancer: A comprehensive review","volume":"15","author":"Kumar","year":"2023","journal-title":"World J. Gastrointest. 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