2. Academic Validation
  3. Nortriptyline induces mitochondria and death receptor-mediated apoptosis in bladder cancer cells and inhibits bladder tumor growth in vivo

Nortriptyline induces mitochondria and death receptor-mediated apoptosis in bladder cancer cells and inhibits bladder tumor growth in vivo

  • Eur J Pharmacol. 2015 Aug 15:761:309-20. doi: 10.1016/j.ejphar.2015.06.007.
Sheau-Yun Yuan 1,  Chen-Li Cheng 2,  Hao-Chung Ho 2,  Shian-Shiang Wang 2,  Kun-Yuan Chiu 2,  Chung-Kuang Su 2,  Yen-Chuan Ou 3,  Chi-Chen Lin 4
Affiliations

Affiliations

  • 1 Division of Urology, Department of Surgery, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC; Department of Nursing, Hung Kung University, Taichung 43302, Taiwan, ROC.
  • 2 Division of Urology, Department of Surgery, Taichung Veterans General Hospital, Taichung 40705, Taiwan, ROC.
  • 3 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: [email protected].
  • 4 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: [email protected].
PMID: 26086857 DOI: 10.1016/j.ejphar.2015.06.007
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.

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).

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