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Regulation of pancreatic cancer growth by superoxide
Journal article   Peer reviewed

Regulation of pancreatic cancer growth by superoxide

Juan Du, Elke S Nelson, Andrean L Simons, Kristen E Olney, Justin C Moser, Hannah E Schrock, Brett A Wagner, Garry R Buettner, Brian J Smith, Melissa L T Teoh, …
Molecular carcinogenesis, Vol.52(7), pp.555-567
07/2013
DOI: 10.1002/mc.21891
PMCID: PMC3375391
PMID: 22392697

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Abstract

K-ras mutations have been identified in up to 95% of pancreatic cancers, implying their critical role in the molecular pathogenesis. Expression of K-ras oncogene in an immortalized human pancreatic ductal epithelial cell line, originally derived from normal pancreas (H6c7), induced the formation of carcinoma in mice. We hypothesized that K-ras oncogene correlates with increased non-mitochondrial-generated superoxide (O 2.-), which could be involved in regulating cell growth contributing to tumor progression. In the H6c7 cell line and its derivatives, H6c7er-Kras+ (H6c7 cells expressing K-ras oncogene), and H6c7eR-KrasT (tumorigenic H6c7 cells expressing K-ras oncogene), there was an increase in hydroethidine fluorescence in cell lines that express K-ras. Western blots and activity assays for the antioxidant enzymes that detoxify O 2.- were similar in these cell lines suggesting that the increase in hydroethidine fluorescence was not due to decreased antioxidant capacity. To determine a possible non-mitochondrial source of the increased levels of O 2.-, Western analysis demonstrated the absence of NADPH oxidase-2 (NOX2) in H6c7 cells but present in the H6c7 cell lines expressing K-ras and other pancreatic cancer cell lines. Inhibition of NOX2 decreased hydroethidine fluorescence and clonogenic survival. Furthermore, in the cell lines with the K-ras oncogene, overexpression of superoxide dismutases that detoxify non-mitochondrial sources of O 2.-, and treatment with the small molecule O 2.- scavenger Tempol, also decreased hydroethidine fluorescence, inhibited clonogenic survival and inhibited growth of tumor xenografts. Thus, O 2.- produced by NOX2 in pancreatic cancer cells with K-ras, may regulate pancreatic cancer cell growth.
Mitochondria - enzymology RNA, Small Interfering - genetics ras Proteins - genetics Proto-Oncogene Proteins p21(ras) Superoxide Dismutase - genetics Cell Proliferation Pancreatic Neoplasms - metabolism Membrane Glycoproteins - metabolism Humans NADPH Oxidases - metabolism Fluorescence Superoxide Dismutase - antagonists & inhibitors ras Proteins - metabolism Spin Labels Cytosol - enzymology Membrane Glycoproteins - antagonists & inhibitors Cyclic N-Oxides Superoxides - metabolism NADPH Oxidases - genetics Tumor Cells, Cultured Superoxide Dismutase - metabolism Proto-Oncogene Proteins - metabolism Tumor Stem Cell Assay NADPH Oxidases - antagonists & inhibitors Pancreatic Neoplasms - pathology Proto-Oncogene Proteins - genetics NADPH Oxidase 2 Blotting, Western Membrane Glycoproteins - genetics Animals Mice, Nude Mice Phenanthridines - metabolism Extracellular Space - enzymology

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