title: Nortriptyline induces mitochondria and death receptor-mediated apoptosis in bladder cancer cells and inhibits bladder tumor growth in vivo.
abstract: Nortriptyline (NTP), an antidepressant, has antitumor effects on some human cancer cells, but its effect on human bladder cancer cells is not known. In this study, we used a cell viability assay to demonstrate that NTP is cytotoxic to human TCCSUP and mouse MBT-2 bladder cancer cells in a concentration and time-dependent manner. We also performed cell cycle analysis, annexin V and mitochondrial membrane potential assays, and Western blot analysis to show that NTP inhibits cell growth in these cells by inducing both mitochondria-mediated and death receptor-mediated apoptosis. Specifically, NTP increases the expression of Fas, FasL, FADD, Bax, Bak, and cleaved forms of caspase-3, caspase-8, caspase-9, and poly(ADP-ribose) polymerase. In addition, NTP decreases the expression of Bcl-2, Bcl-xL, BH3 interacting domain death agonist, X-linked inhibitor of apoptosis protein, and survivin. Furthermore, NTP-induced apoptosis is associated with reactive oxygen species (ROS) production, which can be reduced by antioxidants, such as N-acetyl-L-cysteine. Finally, we showed that NTP suppresses tumor growth in mice inoculated with MBT-2 cells. Collectively, our results suggest that NTP induces both intrinsic and extrinsic apoptosis in human and mouse bladder cancer cells and that it may be a clinically useful chemotherapeutic agent for bladder cancer in humans.
authors.affiliation: Division of Urology, Department of Surgery, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC; Department of Nursing, Hung Kung University, Taichung 43302, Taiwan, ROC.
authors.forename: Sheau-Yun
authors.initials: SY
authors.lastname: Yuan
authors.affiliation: Division of Urology, Department of Surgery, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC.
authors.forename: Chen-Li
authors.initials: CL
authors.lastname: Cheng
authors.affiliation: Division of Urology, Department of Surgery, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC.
authors.forename: Hao-Chung
authors.initials: HC
authors.lastname: Ho
authors.affiliation: Division of Urology, Department of Surgery, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC.
authors.forename: Shian-Shiang
authors.initials: SS
authors.lastname: Wang
authors.affiliation: Division of Urology, Department of Surgery, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC.
authors.forename: Kun-Yuan
authors.initials: KY
authors.lastname: Chiu
authors.affiliation: Division of Urology, Department of Surgery, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC.
authors.forename: Chung-Kuang
authors.initials: CK
authors.lastname: Su
authors.affiliation: Division of Urology, Department of Surgery, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC; Department of Education and Research, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC. Electronic address: ycou@vghtc.gov.tw.
authors.forename: Yen-Chuan
authors.initials: YC
authors.lastname: Ou
authors.affiliation: Institute of Biomedical Science, National Chung-Hsing University, Taichung 40227, Taiwan, ROC; Department of Education and Research, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC. Electronic address: lincc@dragon.nchu.edu.tw.
authors.forename: Chi-Chen
authors.initials: CC
authors.lastname: Lin
mesh_terms: D000818:Animals; D000929:Antidepressive Agents, Tricyclic / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008297:Male; D008809:Mice, Inbred C3H; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D009661:Nortriptyline / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D053218:Receptors, Death Domain / Q000187:drug effects* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D047368:Tumor Burden / Q000187:drug effects*; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology
chemical_list: D000929:Antidepressive Agents, Tricyclic; D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D017382:Reactive Oxygen Species; D053218:Receptors, Death Domain; D009661:Nortriptyline
keywords: 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium Bromide (MTT) (PubChem CID: 64965); Apoptosis; Bladder cancer; DMSO (PubChem CID: 679); Dihydroethidium (DHE) (PubChem CID:128682); Glutathione (GSH) (PubChem CID:124886); JC-1 (PubChem CID:5492929); Mitochondria; N-acetyl-L-cysteine (NAC) (PubChem CID:12035); Nortriptyline; Propidium iodide (PubChem CID:104981); Reactive oxygen species; Tricyclic antidepressant; Tween 20 (PubChem CID:443314)