title: Tetrandrine induces cell death in SAS human oral cancer cells through caspase activation-dependent apoptosis and LC3-I and LC3-II activation-dependent autophagy.
abstract: Numerous studies have demonstrated that autophagy is associated with cancer development. Thus, agents to induce autophagy could be employed in some cases for the treatment of cancer. Our results showed that tetrandrine significantly decreased the viability of SAS cells in a concentration- and time-dependent manner. Tetrandrine induced nuclear condensation, demonstrated by DAPI staining. The early events in apoptosis analysed by Annexin V/PI staining indicated that the percentage of cells staining positive for Annexin V was slightly increased in SAS cells with tetrandrine treatment but was much lower following bafilomycin A1 pre-treatment. Tetrandrine caused AVO and MDC induction in SAS cells in a concentration-dependent manner by fluorescence microscopy. Tetrandrine also caused LC-3 expression in SAS cells in a time-dependent manner. Our results show that tetrandrine treatment induced the levels of cleaved caspase-3 in a concentration- and time-dependent manner. Tetrandrine treatment induced the levels of LC-3 II, Atg-5, beclin-1, p-S6, p-ULK, p-mTOR, p-Akt (S473) and raptor. Tetrandrine decreased cell viability, but bafilomycin A1, 3-MA, chloroquine and NAC protected tetrandrine-treated SAS cells against decrease of cell viability. Atg-5, beclin-1 siRNA decreased tetrandrine-induced cleaved caspase-3 and cleaved PARP in SAS cells and protected tetrandrine-treated SAS cells against decrease in cell viability. Chloroquine, NAC and bafilomycin A1 also decreased tetrandrine-induced cleaved caspase-3 and cleaved PARP in SAS cells. Our results indicate the tetrandrine induces apoptosis and autophagy of SAS human cancer cells via caspase-dependent and LC3-I and LC3-II‑dependent pathways.
authors.affiliation: Department of Nursing, St. Mary's Junior College of Medicine, Nursing and Management, Yilan, Taiwan, ROC.
authors.forename: An-Cheng
authors.initials: AC
authors.lastname: Huang
authors.forename: Jin-Cherng
authors.initials: JC
authors.lastname: Lien
authors.forename: Meng-Wei
authors.initials: MW
authors.lastname: Lin
authors.forename: Jai-Sing
authors.initials: JS
authors.lastname: Yang
authors.forename: Ping-Ping
authors.initials: PP
authors.lastname: Wu
authors.forename: Shu-Jen
authors.initials: SJ
authors.lastname: Chang
authors.forename: Tung-Yuan
authors.initials: TY
authors.lastname: Lai
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D018501:Antirheumatic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics; D001343:Autophagy / Q000187:drug effects*; D000071189:Autophagy-Related Protein-1 Homolog; D000071186:Beclin-1; D044182:Benzylisoquinolines / Q000494:pharmacology*; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002738:Chloroquine / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D018942:Macrolides / Q000494:pharmacology; D008565:Membrane Proteins / Q000235:genetics; D008869:Microtubule-Associated Proteins; D009062:Mouth Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D018501:Antirheumatic Agents; D051017:Apoptosis Regulatory Proteins; C491997:BECN1 protein, human; D000071186:Beclin-1; D044182:Benzylisoquinolines; D004791:Enzyme Inhibitors; D047908:Intracellular Signaling Peptides and Proteins; C477012:MAP1LC3A protein, human; D018942:Macrolides; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; C009438:tetrandrine; D002738:Chloroquine; C040929:bafilomycin A1; D011065:Poly(ADP-ribose) Polymerases; D000071189:Autophagy-Related Protein-1 Homolog; D017346:Protein Serine-Threonine Kinases; C484754:ULK1 protein, human; D053148:Caspase 3