Metastatic suppressor CD44 is related with oxidative stress in breast cancer cell lines

Evidence has accumulated on the role of reactive oxygen species (ROS) in metastasis since surgical removal of tumors generates oxidative stress promoting metastasis and cell growth. Metastasis consists of a cascade of events which allow the cell to survive in target tissues and influence several processes such as dissemination from tumor tissue, transport in blood/lymphatic vessels, invasion and homing of malignant cells. A cDNA oligoarray was used to determine whether alterations of metastatic genes are associated with oxidative stress in breast cancer cell lines. The cell lines used for the experiments were derived from a pre-existent in vitro breast cancer progression model originated in our laboratory. The cDNA array showed alterations in functional gene groups related with cell-cell and cell-matrix interaction molecules, such as caveolin-1; metastasis suppressor genes, such as CD44; metastasis-associated proteases, such as cathepsin D and the protease inhibitor, plasminogen activator inhibitor type 1. The changes of the selected genes were validated by differential display-RT-PCR as well as by protein expression assessed by Western blot analysis. It was found that the cell line, called Tumor2 with down-regulation of basal ROS and manganese superoxide dismutase (MnSOD) expression as a constitutive pattern of this cell line, presented alterations in genes that confer metastatic potential in comparison to the Alpha5 cell line, showing overexpression of basal MnSOD and high levels of ROS. Interesting, it was to found that CD44, considered a metastatic suppressor gene, was influenced by ROS, measured by hydrogen peroxide treatments, as seen by decreased CD44 protein expression in the Alpha5 cell line in a compensatory response to increased MnSOD protein expression. In conclusion, alterations of metastatic genes in malignant breast cancer cell lines were observed in relation to ROS and basal levels of antioxidant enzymes.