MnSOD inhibits proline oxidase-induced apoptosis in colorectal cancer cells

• Published in: Carcinogenesis (New York) Vol. 26; no. 8; pp. 1335 - 1342
• Main Authors: Liu, Yongmin, Borchert, Gregory L., Donald, Steven P., Surazynski, Arkadiusz, Hu, Chien-An, Weydert, Christine J., Oberley, Larry W., Phang, James M.
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01.08.2005; Oxford Publishing Limited (England)
• Subjects: 7′-dichloro-fluorescein diacetate; Acetylcysteine - pharmacology; adenovirus; Apoptosis; Biological and medical sciences; Carcinogenesis, carcinogens and anticarcinogens; CAT; catalase; Catalase - metabolism; Cell Death - drug effects; Cell Line, Tumor; Colorectal cancer; Colorectal Neoplasms; copper-zinc superoxide dismutase; CuZnSOD; DCFH; DOX; doxycycline; Humans; Hydrogen Peroxide - metabolism; Kinetics; manganese superoxide dismutase; Medical sciences; MnSOD; MOI; multiplicity of infection; N-acetyl cysteine; NAC; p53-induced gene; P5C; PARP; PBS; phosphate-buffered saline; PIG; poly (ADP-ribose) polymerase; POX; proline oxidase; Proline Oxidase - metabolism; pyrroline-5-carboxylate; reactive nitrogen species; Reactive Oxygen Species; Recombinant Proteins - metabolism; RNS; ROS; Superoxide Dismutase - metabolism; Transfection; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Tumors; Rectal disease; Enzyme; Colorectal cancer; Oxidase; Proline; Superoxide dismutase; Malignant tumor; Colonic disease; Cell death; Aminoacid; Digestive diseases; Intestinal disease; Inhibitor; Oxidoreductases; Tumor cell; Apoptosis
• ISSN: 0143-3334; 1460-2180
• DOI: 10.1093/carcin/bgi083

Proline oxidase (POX), localized on inner mitochondrial membranes, is encoded by a p53-induced gene and metabolically participates in p53-induced apoptosis. Previously, we showed that POX catalyzed the generation of reactive oxygen species (ROS). We and others have demonstrated that overexpression of POX, independent of p53, causes apoptotic cell death in a variety of cancer cells. But a necessary role for ROS remains uncertain. Therefore, we asked whether superoxide dismutases (SOD) and catalase (CAT), important antioxidant enzymes, might interfere with the POX-dependent induction of apoptosis. In this study, we used DLD-1 colorectal cancer cells stably transfected with the POX gene under the control of a tetracycline-inducible promoter. When doxycycline was removed from the culture medium and the expression of POX was induced, apoptotic cell death was initiated. To examine the importance of the ROS-dependent component of the pathway, we infected DLD-1 POX cells with recombinant adenoviruses containing MnSOD, CuZnSOD, CAT or varying combinations of these adenoviruses followed by induced expression of POX. The expression of MnSOD inhibited POX-induced apoptosis, but others did not. Mechanistically, mitochondria-localized MnSOD dramatically reduced the release of cytochrome c to cytosol by POX. Compared with control cells, MnSOD-expressing DLD-1 POX cells generated a higher concentration of H2O2 owing to dismutation of superoxide radicals, which was elevated by POX. Thus, these data further suggest that the generation of superoxide radicals plays a crucial role in POX-induced apoptosis and the process is partially blocked by MnSOD.