Timosaponin AIII is preferentially cytotoxic to tumor cells through inhibition of mTOR and induction of ER stress

• Published in: PloS one Vol. 4; no. 9; p. e7283
• Main Authors: King, Frank W, Fong, Sylvia, Griffin, Chandi, Shoemaker, Mark, Staub, Rick, Zhang, Yan-Ling, Cohen, Isaac, Shtivelman, Emma
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 30.09.2009; Public Library of Science (PLoS)
• Subjects: Analysis; Anemarrhena - metabolism; Apoptosis; Autophagy; Breast cancer; Cancer; Cancer therapies; Caspase; Caspase-4; Cell Biology/Cellular Death and Stress Responses; Cell culture; Cell cycle; Cell death; Cell growth; Cell Line, Transformed; Cell Line, Tumor; Chemical compounds; Chemical fractionation; Cholesterol; Cytotoxicity; Development and progression; Drug Screening Assays, Antitumor; Endoplasmic reticulum; Endoplasmic Reticulum - metabolism; Flow Cytometry; Fractionation; Gene expression; Gene Expression Regulation, Neoplastic; Glycosylation; Homeostasis; Humans; Inhibition; Kinases; Molecular Biology; Mortality; Oncology/Breast Cancer; Pathways; Phagocytosis; Pharmacology; Phosphorylation; Plant Extracts - pharmacology; Protein Kinases - metabolism; Proteins; Quality control; Restoration; Saponins; Saponins - pharmacology; Selectivity; Signaling; Steroids - pharmacology; Sterols; Stress response; Stresses; Structure-Activity Relationship; Sugar; TOR protein; TOR Serine-Threonine Kinases; Toxicity; Transformed cells; Tumor cell lines; Tumor cells; Tumors; United States > US; California
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0007283

The aqueous extract of Anemarrhena asphodeloides (BN108) induces apoptosis in various cancer cell lines but is significantly less cytotoxic in non-transformed cells. Chemical fractionation of BN108 showed that its cytotoxicity is associated with timosaponins, steroidal saponins of coprostane type. Timosaponin BII (TBII) is a major saponin in BN108, but it shows little cytotoxicity. A much less abundant TAIII induces cell death in tumor cells but not in normal cells, reproducing the selectivity of the total extract BN108. Glycosidase treatment, by removing the extra sugar moiety in TBII, converts it to TAIII and confers cytotoxic activity. Analysis of the mechanisms of death induced by TAIII revealed activation of two distinct pro-apoptotic pathways: first, inhibition of mTORC1 manifested in much reduced phosphorylation of mTORC1 targets; second, induction of endoplasmic reticulum stress culminating in phosphorylation of eIF2alpha and activation of caspase 4. These pro-apoptotic pathways are activated by TAIII selectively in tumor cells but not in normal cells. Both pathways play a causative role in TAIII cytotoxicity, as restoration of either mTOR activity or relief of ER stress alone offer only partial protection from TAIII. Inhibition of mTORC1 and induction of ER stress apparently contribute to the induction of the previously reported autophagic response in TAIII-treated cells. TAIII induced autophagy plays a protective role in TAIII induced death signaling, and failure to mount autophagic response is associated with heightened sensitivity to TAIII induced apoptosis. The multiple death-promoting and apparently tumor-selective responses to TAIII, its ability to inhibit mTORC1, and the possibility of further enhancing its cytotoxicity by pharmacological inhibition of autophagy, make TAIII an attractive candidate for development as a cancer therapeutic agent.