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This study determines the role of laminin-1 in promoting metastatic colonization during breast cancer. For this purpose, human mammary epithelial cell lines representing normal (MCF-10A), adenocarcinoma (MCF-7), and malignant carcinoma (MDA-MB-231) were propagated in 3-dimensional cultures composed of laminin-1, collagen I, or mixtures of the two, and analyzed by Western blot, immunocytochemistry, semiquantitative reverse transcription polymerase chain reaction, and methylation-specific PCR. Here we demonstrate that laminin-1 decreases methylation of the E-cadherin promoter, resulting in increased mRNA and protein expression for malignant mammary epithelial cells. This decreased methylation is associated with dramatic changes in the cellular and structural morphology as well as a 70-fold decrease in DNA methyltransferase 1 (DNMT1) and a 6-fold decrease in cadherin 11 protein expression. To control for specificity of laminin-1 interactions, cells were also cultured on 2-dimensional plastic substrata and collagen I hydrogels for analysis, and the MCF-10A and MCF-7 were used as nonmalignant controls. Using a 3-dimensional model, we present evidence that laminin-1 is capable of inducing epigenetic change by inhibiting expression of DNMT1 and preventing methylation of the E-cadherin promoter, resulting in E-cadherin expression and the formation of cell-cell bonds in malignant carcinoma.—Benton, G., Crooke, E., George, J. Laminin-1 induces E-cadherin expression in 3-dimensional-cultured breast cancer cells by inhibiting DNA methyltransferase 1 and reversing promoter methylation status.
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