Regulation of Id-1 Gene Expression in Aggressive Breast Cancer and Glioblastoma Cells Using Cannabinoid Compounds
AbstractWe investigated Id-1 gene expression in aggressive breast cancer cells and glioblastoma cells, and compared Id-1 expression in control cells with cells treated with one of the cannabinoid compounds, cannabidiol (CBD) or tetrahydrocannabinol (THC). The tumor cells were treated with CBD or THC, and the control group was treated with ethanol. After 2-3 days of treatment, cells were harvested and proteins extracted from them. Following protein electrophoresis, Western blot technique was used to analyse the expression of Id-1 protein derived from the samples. Using a rabbit monoclonal anti-human Id-1 antibody, and a goat anti-rabbit HRP as a secondary antibody, the intensities of the bands produced following film exposure were used to compare drug-treated cells with controls. Id-1 gene expression was decreased in tumor cells treated with cannabinoid compounds. We also aimed to visualize tumor cell migration through a living tissue and to determine the possible inhibition of migration upon treatment with CBD. Tumor cell migration through a living tissue was studied by placing GFP-labelled glioblastoma cells on top of Sprague-Dawley rat brain slices. Migration was studied using the Carl-Zeis cell monitor. Glioblastoma cells placed on brain slices of Sprague-Dawley rats were found to migrate through the tissue to the other side of the slice. The morphology of glioblastoma cells was also found to be modified upon treatment (changes in cell-to-cell contacts). This study shows that Id-1 downregulation strongly correlates with a reduced tumor aggressiveness and that Id-1 might represent a potent therapeutic target for breast cancer and glioblastoma.
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