Invasive potential induced under long-term oxidative stress in mammary epithelial cells.
Although the causal relationship between chronic inflammation and carcinogenesis has long been discussed, the molecular basis of the relation is poorly understood. In the present study, we focused on reactive oxygen species (ROS) and their signals under inflammatory conditions leading to the carcinogenesis of epithelial cells and found that repeated treatment with a low dose of H(2)O(2) (0.2 mmol/L) for periods of 2 to 4 days caused a phenotypic conversion of mouse NMuMG mammary epithelial cells from epithelial to fibroblast-like as in malignant transformation. The phenotypic conversion included the dissolution of cell-cell contacts, redistribution of E-cadherin in the cytoplasm, and up-regulation of a set of integrin family members (integrin alpha2, alpha6, and beta3) and matrix metalloproteinases (MMPs; MMP-3, -10, and -13), as analyzed using Northern blot analysis and quantitative reverse transcription-PCR. Gelatin zymography indicated post-transcriptional activation of gelatinases, including MMP-2 and -9. In parallel, p38 mitogen-activated protein kinase and extracellular signal-regulated kinase 1/2 were activated, which contributed to the induction of MMP-13, and a glutathione S-transferase pull-down assay showed the activation of a small GTPase, Rac1. Surprisingly, the prolonged oxidative treatment was sufficient to induce all of the aforementioned events. Most importantly, depending on the MMP activities, the epithelial cells exposed to oxidative conditions eventually acquired invasiveness in a reconstituted model system with a Matrigel invasion chamber containing normal fibroblasts at the bottom, providing the first substantial evidence supporting the direct role of ROS signals in the malignant transformation of epithelial cells.
Mori
Kazunori
K
Department of Microbiology, Showa University School of Pharmaceutical Sciences, Tokyo, Japan.
Shibanuma
Motoko
M
Nose
Kiyoshi
K
D000818:Animals; D002471:Cell Transformation, Neoplastic / Q000378:metabolism*; D017364:Collagenases / Q000096:biosynthesis; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology; D007527:Isoenzymes; D008321:Mammary Glands, Animal / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D008325:Mammary Neoplasms, Experimental / Q000378:metabolism* / Q000473:pathology; D053509:Matrix Metalloproteinase 13; D020782:Matrix Metalloproteinases / Q000096:biosynthesis / Q000378:metabolism; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009479:Neuropeptides / Q000096:biosynthesis / Q000235:genetics; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015854:Up-Regulation; D020744:rac GTP-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D020830:rac1 GTP-Binding Protein
D007527:Isoenzymes; D009479:Neuropeptides; C484602:Rac1 protein, mouse; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D020928:Mitogen-Activated Protein Kinases; D017364:Collagenases; D053509:Matrix Metalloproteinase 13; D020782:Matrix Metalloproteinases; C507081:Mmp13 protein, mouse; D020744:rac GTP-Binding Proteins; D020830:rac1 GTP-Binding Protein