PloS one, 2020-05, Vol.15 (5), p.e0233208-e0233208
2020
Details
- Autor(en) / Beteiligte
- Titel
- NADPH oxidase 1 is highly expressed in human large and small bowel cancers
- Ist Teil von
- PloS one, 2020-05, Vol.15 (5), p.e0233208-e0233208
- Ort / Verlag
- United States: Public Library of Science
- Erscheinungsjahr
- 2020
- Link zum Volltext
- Quelle
- Electronic Journals Library - Freely accessible e-journals
- Beschreibungen/Notizen
- To facilitate functional investigation of the role of NADPH oxidase 1 (NOX1) and associated reactive oxygen species in cancer cell signaling, we report herein the development and characterization of a novel mouse monoclonal antibody that specifically recognizes the C-terminal region of the NOX1 protein. The antibody was validated in stable NOX1 overexpression and knockout systems, and demonstrates wide applicability for Western blot analysis, confocal microscopy, flow cytometry, and immunohistochemistry. We employed our NOX1 antibody to characterize NOX1 expression in a panel of 30 human colorectal cancer cell lines, and correlated protein expression with NOX1 mRNA expression and superoxide production in a subset of these cells. Although a significant correlation between oncogenic RAS status and NOX1 mRNA levels could not be demonstrated in colon cancer cell lines, RAS mutational status did correlate with NOX1 expression in human colon cancer surgical specimens. Immunohistochemical analysis of a comprehensive set of tissue microarrays comprising over 1,200 formalin-fixed, paraffin-embedded tissue cores from human epithelial tumors and inflammatory disease confirmed that NOX1 is overexpressed in human colon and small intestinal adenocarcinomas, as well as adenomatous polyps, compared to adjacent, uninvolved intestinal mucosae. In contradistinction to prior studies, we did not find evidence of NOX1 overexpression at the protein level in tumors versus histologically normal tissues in prostate, lung, ovarian, or breast carcinomas. This study constitutes the most comprehensive histopathological characterization of NOX1 to date in cellular models of colon cancer and in normal and malignant human tissues using a thoroughly evaluated monoclonal antibody. It also further establishes NOX1 as a clinically relevant therapeutic target in colorectal and small intestinal cancer.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1932-6203eISSN: 1932-6203DOI: 10.1371/journal.pone.0233208Titel-ID: cdi_plos_journals_2404631257
- Format
- –
- Schlagworte
- Adenocarcinoma - enzymology, Adenocarcinoma - genetics, Adenocarcinoma - pathology, Antibodies, Biology and Life Sciences, Biotechnology, Breast, Breast cancer, Breast carcinoma, Caco-2 Cells, Cancer, Cancer research, Cancer therapies, Cell culture, Cellular signal transduction, Characterization, Cloning, Colon, Colon cancer, Colonic Neoplasms - enzymology, Colonic Neoplasms - genetics, Colorectal cancer, Colorectal carcinoma, Confocal microscopy, Correlation, Development and progression, Diseases, Epithelial tumors, Flow cytometry, Formaldehyde, Gastrointestinal cancer, Gene expression, Gene Expression Regulation, Enzymologic, Gene Expression Regulation, Neoplastic, Genetic aspects, Health aspects, HT29 Cells, Human tissues, Humans, Immunohistochemistry, Inflammation, Inflammatory diseases, Intestinal cancer, Intestine, Intestine, Small - enzymology, Intestine, Small - pathology, Laboratories, Lung carcinoma, Medical research, Medicine and Health Sciences, Membrane proteins, Messenger RNA, Microscopy, Models, Biological, Monoclonal antibodies, NAD(P)H oxidase, NADPH Oxidase 1 - biosynthesis, NADPH Oxidase 1 - genetics, Neoplasm Proteins - biosynthesis, Neoplasm Proteins - genetics, Novels, Oncology, Experimental, Ovarian carcinoma, Oxidase, Oxidases, Oxygen, Paraffin, Paraffins, Physical Sciences, Polyps, Prostate, Proteins, Ras protein, Reactive oxygen species, Research and Analysis Methods, Respiration, RNA, Small intestine, Superoxide, Superoxides, Therapeutic applications, Tissues, Tumor cell lines, Tumors