{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T14:13:53Z","timestamp":1778249633274,"version":"3.51.4"},"reference-count":53,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T00:00:00Z","timestamp":1777248000000},"content-version":"vor","delay-in-days":26,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T00:00:00Z","timestamp":1775001600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cell Biochemistry &amp;amp; Function"],"published-print":{"date-parts":[[2026,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Cytochrome C (Cyt C) is a central mediator of intrinsic apoptosis, whereas its heme\u2010free precursor, apocytochrome C (APO\u2010Cyt C), competitively inhibits this process. Sodium aescinate (SA), a natural triterpene saponin, is known to facilitate the endosomal escape of protein drugs. We initially aimed to investigate the effects of enhancing endosomal escape efficiency on protein activity through the combination of SA and Cyt C. However, this study investigates an cytotoxicity observed when APO\u2010Cyt C is combined with SA and aims to elucidate the underlying molecular mechanism. Contrary to its established anti\u2010apoptotic function, APO\u2010Cyt C, when co\u2010administered with a non\u2010toxic concentration of SA, induced potent, caspase\u2010dependent mitochondrial apoptosis in cancer cells. This pro\u2010apoptotic switch was not primarily triggered by BCL\u20102 family protein modulation, ROS generation, or calcium overload. Instead, the primary mechanism is the induction of excessive and lethal autophagy. SA was found to induce lysosomal membrane damage, evidenced by Galectin\u20109 recruitment, which initiates lysophagy. The addition of APO\u2010Cyt C significantly amplified the autophagic flux, leading to decreased p62 levels and enhanced LC3\u2010II turnover. Mechanistically, this synergy is driven by a dual impact on the AKT\u2010mTOR\u2010TFEB pathway: APO\u2010Cyt C treatment decreased mTOR phosphorylation, while the combination promoted the nuclear translocation of the autophagy regulator, TFEB. Inhibition of autophagic flux using Bafilomycin A1 or Tetrandrine rescued cells from apoptosis, confirming that excessive autophagy is the direct cause of cell death. This study reveals a novel therapeutic strategy wherein an anti\u2010apoptotic protein is converted into a potent pro\u2010apoptotic agent. The combination of APO\u2010Cyt C and SA triggers apoptosis by overwhelming the cell with excessive autophagic flux, driven by synergistic inhibition of the mTOR\u2010TFEB axis. These findings highlight the therapeutic potential of modulating autophagy and suggest that combining mTOR inhibitors with lysosome\u2010targeting agents like SA could be an effective anti\u2010cancer strategy.<\/jats:p>","DOI":"10.1002\/cbf.70214","type":"journal-article","created":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T04:56:52Z","timestamp":1777352212000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Synergistic Induction of Apoptosis by Apocytochrome C and Sodium Aescinate Is Mediated by Excessive Autophagy via the AKT\u2010mTOR\u2010TFEB Signaling Axis"],"prefix":"10.1002","volume":"44","author":[{"given":"Jia\u2010Wei","family":"Sun","sequence":"first","affiliation":[{"name":"Department of Applied Biology East China University of Science and Technology Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yin\u2010Kang","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Applied Biology East China University of Science and Technology Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Liang","family":"Ji","sequence":"additional","affiliation":[{"name":"Department of Applied Biology East China University of Science and Technology Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jian","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Applied Biology East China University of Science and Technology Shanghai China"},{"name":"ECUST\u2010FONOW Joint Research Center for Innovative Medicines East China University of Science and Technology Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fu\u2010Jun","family":"Wang","sequence":"additional","affiliation":[{"name":"ECUST\u2010FONOW Joint Research Center for Innovative Medicines East China University of Science and Technology Shanghai China"},{"name":"New Drug R&amp;D Center Zhejiang Fonow Medicine Co. 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