Premium
Ouabain induces apoptotic cell death in human prostate DU 145 cancer cells through DNA damage and TRAIL pathways
Author(s) -
Chang YiMing,
Shih YungLuen,
Chen ChaoPing,
Liu KoLin,
Lee MeiHui,
Lee MingZhe,
Hou HsinTu,
Huang HsiehChou,
Lu HsuFeng,
Peng ShuFen,
Chen KuoWei,
Yeh MingYang,
Chung JingGung
Publication year - 2019
Publication title -
environmental toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.813
H-Index - 77
eISSN - 1522-7278
pISSN - 1520-4081
DOI - 10.1002/tox.22834
Subject(s) - apoptosis , ouabain , viability assay , microbiology and biotechnology , fadd , dna damage , fas ligand , cancer cell , biology , programmed cell death , cell cycle , chemistry , cytochrome c , caspase , biochemistry , cancer , dna , genetics , organic chemistry , sodium
Ouabain, a cardiotonic steroid and specific Na + /K + ‐ATPase inhibitor, has a potential to induce cancer cell apoptosis but the mechanisms of apoptosis induced by ouabain are not fully understand. The aim of this study was to investigate the cytotoxic effects of ouabain on human prostate cancer DU 145 cells in vitro. Cell morphological changes were examined by phase contrast microscopy. Cell viability, cell cycle distribution, cell apoptosis, DNA damage, the production of ROS and Ca 2+ , and mitochondrial membrane potential (ΔΨ m ) were measured by flow cytometry assay. Results indicated that ouabain induced cell morphological changes, decreased total cell viability, induced G0/G1 phase arrest, DNA damage, and cell apoptosis, increased ROS and Ca 2+ production, but decreased the levels of ΔΨ m in DU 145 cells. Ouabain also increased the activities of caspase‐3, ‐8, and ‐9. Western blotting was used for measuring the alterations of apoptosis‐associated protein expressions in DU 145 cells and results indicated that ouabain increased the expression of DNA damage associated proteins (pATM Ser1981 , p‐H2A.X Ser139 , and p‐p53 Ser15 ) and ER‐stress‐associated proteins (Grp78, ATF6β, p‐PERK Thr981 , PERK, eIF2A, GADD153, CaMKIIβ, and caspase‐4) in time‐dependently. Furthermore, ouabain increased apoptosis‐associated proteins (DR4, DR5, Fas, Fas Ligand, and FADD), TRAIL pathway, which related to extrinsic pathway, promoted the pro‐apoptotic protein Bax, increased apoptotic‐associated proteins, such as cytochrome c, AIF, Endo G, caspase‐3, ‐8, and ‐9, but reduced anti‐apoptotic protein Bcl‐2 and Bcl‐x in DU 145 cells. In conclusion, we may suggest that ouabain decreased cell viability and induced apoptotic cell death may via caspase‐dependent and mitochondria‐dependent pathways in human prostate cancer DU 145 cells.