Characterization of NADPH oxidase 5 expression in human tumors and tumor cell lines with a novel mouse monoclonal antibody |
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| Institution: | 1. Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA;2. Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA;3. Division of Cancer Treatment and Diagnosis, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA;4. Pathology/Histotechnology Laboratory, SAIC Frederick, Inc./Frederick National Laboratory for Cancer Research, National Institutes of Health, Frederick, MD 21702, USA;1. Cancer Genetics Lab, Department of Medical and Molecular Genetics, King''s College London School of Medicine, 8th Floor, Tower Wing, Guy''s Hospital, London SE1 9RT, UK;2. Department of Haematology, Cardiff University, Cardiff, UK;1. Laboratory of Pharmacology and Pathophysiology, Faculty of Pharmacy, Kindai University (formerly known as Kinki University), Higashi-Osaka, 577-8502, Japan;2. Molecular Regulation of Aging, Tokyo Metropolitan Institute of Gerontology, Tokyo, 173-0015, Japan;1. Laboratory of Thermodynamics of Solutions of Non-electrolytes and Biologically Active Substances, G.A. Krestov Institute of Solution Chemistry, Russian Academy of Sciences, 1 Akademicheskaya Str., 153045 Ivanovo, Russian Federation;2. Incorporated Physicochemical Center of Solution Researches, G.A. Krestov Institute of Solution Chemistry, Russian Academy of Sciences, 1 Akademicheskaya Str., 153045 Ivanovo, Russian Federation;3. Department of Inorganic Chemistry, Ivanovo’s State University of Chemistry and Technology, 7 Sheremetevsky Ave., 153000 Ivanovo, Russian Federation;4. Laboratory of Nitrogen-containing Compounds, N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 47 Leninsky Ave., 199119 Moscow, Russian Federation;1. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Rd., Qingdao, Shandong 266071, China;2. University of Chinese Academy of Sciences, Beijing 100049, China;1. State Key Laboratory of Ultrasound Engineering in Medicine Co-Founded by Chongqing and The Ministry of Science and Technology, Chongqing Key Laboratory of Biomedical Engineering, College of Biomedical Engineering, Chongqing Medical University, Chongqing 400016, PR China;2. Institute of Life Science, Chongqing Medical University, Chongqing 400016, PR China;3. Department of Obstetrics and Gynecology, Chongqing Hifu Hospital, Chongqing 401121, PR China |
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| Abstract: | Reactive oxygen species generated by NADPH oxidase 5 (Nox5) have been implicated in physiological and pathophysiological signaling pathways, including cancer development and progression. However, because immunological tools are lacking, knowledge of the role of Nox5 in tumor biology has been limited; the expression of Nox5 protein across tumors and normal tissues is essentially unknown. Here, we report the characterization and use of a mouse monoclonal antibody against a recombinant Nox5 protein (bp 600–746) for expression profiling of Nox5 in human tumors by tissue microarray analysis. Using our novel antibody, we also report the detection of endogenous Nox5 protein in human UACC-257 melanoma cells. Immunofluorescence, confocal microscopy, and immunohistochemical techniques were employed to demonstrate Nox5 localization throughout UACC-257 cells, with perinuclear enhancement. Tissue microarray analysis revealed, for the first time, substantial Nox5 overexpression in several human cancers, including those of prostate, breast, colon, lung, brain, and ovary, as well as in malignant melanoma and non-Hodgkin lymphoma; expression in most nonmalignant tissues was negative to weak. This validated mouse monoclonal antibody will promote further exploration of the functional significance of Nox5 in human pathophysiology, including tumor cell growth and proliferation. |
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| Keywords: | NADPH oxidase 5 Tissue microarray Mouse monoclonal Nox5 antibody Melanoma UACC-257 cells Reactive oxygen species Superoxide Free radicals |
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