{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T19:37:10Z","timestamp":1786390630472,"version":"build-2736575974"},"reference-count":53,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2026,4,6]],"date-time":"2026-04-06T00:00:00Z","timestamp":1775433600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,4,6]],"date-time":"2026-04-06T00:00:00Z","timestamp":1775433600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Vet Comparative Oncology"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Canine pulmonary adenocarcinoma (cPAC) lacks well\u2010validated systemic treatment options, motivating preclinical identification of potential therapeutic candidates. To explore potential therapeutic candidates, we first performed a screening of an FDA\u2010approved drug library using a cPAC cell line, which identified bortezomib\u2014a proteasome inhibitor\u2014as a potential agent. Guided by this result, we focused subsequent investigations on proteasome blockade and evaluated the antitumor activity and mechanisms of bortezomib together with a second\u2010generation proteasome inhibitor, carfilzomib\u2014alone and in combination with standard cytotoxics\u2014in three cPAC cell lines (CLAC, HDC, and LuBi). Both agents reduced cell proliferation and cell viability in a dose\u2010dependent manner, with proteasome target engagement (ubiquitinated protein accumulation), G2\/M arrest, sub\u2010G1 increase, and caspase\u20103 cleavage. Pan\u2010 and selective caspase inhibition partially rescued viability, suggesting contributions from both intrinsic and extrinsic apoptosis, together with caspase\u2010independent mechanisms. We also observed activation of the JNK\u2013c\u2010Jun pathway and increased Bim expression, indicating the induction of cellular stress responses and apoptosis. Drug\u2010interaction analysis showed reproducible synergy between carfilzomib and carboplatin in HDC and LuBi, whereas combinations with vinorelbine were mainly additive or antagonistic effects. Overall, proteasome inhibitors exert multifaceted cytotoxic effects in cPAC through proteotoxic stress, cell\u2010cycle arrest, and caspase\u2010associated apoptosis. In particular, the combination of carfilzomib and carboplatin may represent a promising therapeutic strategy. Further in\u00a0vivo and clinical investigations are warranted to evaluate therapeutic efficacy, safety, and pharmacokinetics of proteasome inhibitors in cPAC.<\/jats:p>","DOI":"10.1111\/vco.70065","type":"journal-article","created":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T04:36:21Z","timestamp":1775536581000},"page":"421-432","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Proteasome Inhibitor\u2013Induced Cytotoxic Mechanisms in Canine Pulmonary Adenocarcinoma Cell Lines"],"prefix":"10.1111","volume":"24","author":[{"given":"Aika","family":"Chijiwa","sequence":"first","affiliation":[{"name":"Laboratory of Molecular Diagnostics and Therapeutics, Joint Faculty of Veterinary Medicine Yamaguchi University  Yamaguchi Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sakuya","family":"Inanaga","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Diagnostics and Therapeutics, Joint Graduate School of Veterinary Medicine Yamaguchi University  Yamaguchi Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomohiro","family":"Osaki","sequence":"additional","affiliation":[{"name":"Joint Department of Veterinary Medicine, Faculty of Agriculture Tottori University  Tottori Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masaya","family":"Igase","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Diagnostics and Therapeutics, Joint Faculty of Veterinary Medicine Yamaguchi University  Yamaguchi Japan"},{"name":"Japan Small Animal Cancer Center Public Interest Incorporated Foundation Japan Small Animal Medical Center  Saitama Japan"},{"name":"Division of Translational Research for One Medicine, Research Institute for Cell Design Medical Science Yamaguchi University  Yamaguchi Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takuya","family":"Mizuno","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Diagnostics and Therapeutics, Joint Faculty of Veterinary Medicine Yamaguchi University  Yamaguchi Japan"},{"name":"Japan Small Animal Cancer Center Public Interest Incorporated Foundation Japan Small Animal Medical Center  Saitama Japan"},{"name":"Division of Translational Research for One Medicine, Research Institute for Cell Design Medical Science Yamaguchi University  Yamaguchi Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,6]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.2460\/javma.1989.195.01.106"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1177\/030098589603300601"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1177\/030098588101800409"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.2460\/javma.20.12.0698"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2020.519703"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/jvim.16739"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1939-1676.2004.tb02581.x"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.3390\/vetsci10120664"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/vco.13069"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/vco.12622"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0962-8924(98)01346-4"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/349132a0"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7600808"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.26.5.1967-1978.2006"},{"issue":"11","key":"e_1_2_12_16_1","first-page":"2615","article-title":"Proteasome Inhibitors: A Novel Class of Potent and Effective Antitumor Agents","volume":"59","author":"Adams J.","year":"1999","journal-title":"Cancer Research"},{"issue":"7","key":"e_1_2_12_17_1","first-page":"3071","article-title":"The Proteasome Inhibitor PS\u2010341 Inhibits Growth, Induces Apoptosis, and Overcomes Drug Resistance in Human Multiple Myeloma Cells","volume":"61","author":"Hideshima T.","year":"2001","journal-title":"Cancer Research"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2007-01-065888"},{"issue":"3","key":"e_1_2_12_19_1","first-page":"1145","article-title":"Mechanisms of Proteasome Inhibitor PS\u2010341\u2010Induced G(2)\u2010M\u2010Phase Arrest and Apoptosis in Human Non\u2010Small Cell Lung Cancer Cell Lines","volume":"9","author":"Ling Y. 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