title: Functional activity and tumor-specific expression of dual oxidase 2 in pancreatic cancer cells and human malignancies characterized with a novel monoclonal antibody.
abstract: Dual oxidase 2 (Duox2), one of the seven members of the NADPH oxidase gene family, plays a critical role in generating H2O2 for thyroid hormone biosynthesis and as an integral part of the host defense system of the respiratory epithelium and the gastrointestinal tract. Recent evidence suggests that the regulation of Duox2 expression is under the control of pro-inflammatory cytokines and that Duox2-induced reactive oxygen species (ROS) contribute to the inflammation-related tissue injury that occurs in two pre-malignant, inflammatory conditions: chronic pancreatitis and inflammatory bowel disease. Because no reliable Duox antibodies are commercially available, we report the development of a murine monoclonal antibody (MAb) to Duox2 (clone Duox S-12) and its use for the characterization of Duox2 expression in human tumors, tumor cell lines and normal tissues. Duox S-12 specifically detected both endogenously- and ectopically-expressed Duox2 protein by immunoblotting, immunofluorescence microscopy and immunohistochemistry (where both membranous and cytoplasmic staining were present). Duox2 expression detected by Duox S-12 was functionally coupled to the generation of H(2)O(2) in pancreatic cancer cells that expressed Duox2 and its cognate maturation factor DuoxA2. Although Duox S-12 recognizes ectopically expressed Duox1 protein because of the extensive amino acid homology between Duox1 and Duox2, the lack of substantial Duox1 mRNA expression in human tumors (except thyroid cancer) allowed us to evaluate Duox2 expression across a wide range of normal and malignant tissues by immuno-histochemistry. Duox2 was expressed at elevated levels in many human cancers, most notably tumors of the prostate, lung, colon and breast while brain tumors and lymphomas demonstrated the lowest frequency of expression. The Duox-specific monoclonal antibody described here provides a promising tool for the further examination of the role of Duox-dependent reactive oxygen production in inflammation-related carcinogenesis, where alterations in oxidant tone play a critical role in cell growth and proliferation.
authors: Laboratory of Molecular Pharmacology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors: Yonghzong
authors: Y
authors: Wu
authors: Smitha
authors: S
authors: Antony
authors: Stephen M
authors: SM
authors: Hewitt
authors: Guojian
authors: G
authors: Jiang
authors: Sherry X
authors: SX
authors: Yang
authors: Jennifer L
authors: JL
authors: Meitzler
authors: Agnes
authors: A
authors: Juhasz
authors: Jiamo
authors: J
authors: Lu
authors: Han
authors: H
authors: Liu
authors: James H
authors: JH
authors: Doroshow
authors: Krishnendu
authors: K
authors: Roy
mesh_terms: D000230:Adenocarcinoma / Q000201:enzymology*; D000595:Amino Acid Sequence; D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry / Q000276:immunology*; D000918:Antibody Specificity; D001943:Breast Neoplasms / Q000201:enzymology*; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D000074623:Dual Oxidases; D005260:Female; D006801:Humans; D006825:Hybridomas; D006861:Hydrogen Peroxide / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019255:NADPH Oxidases / Q000276:immunology / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000201:enzymology*; D046888:Tissue Array Analysis
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; C513411:DUOXA2 protein, human; D008565:Membrane Proteins; D006861:Hydrogen Peroxide; D000074623:Dual Oxidases; D019255:NADPH Oxidases; C431515:DUOX2 protein, human
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