Selection of a MCF-7 Breast Cancer Cell Subpopulation with High Sensitivity to IL-1β: Characterization of and Correlation between Morphological and Molecular Changes Leading to Increased Invasiveness
Figure 4
Characterization of E-cadherin mRNA expression and E-cadherin levels in MCF-7pl and MCF-7A3 cells. (A) Relative expression profile of E-cadherin mRNA. MCF7-7pl and MCF-7A3 cells were stimulated with IL-1β as previously indicated. After stimulus, gene expression was evaluated by qRT-PCR at the indicated time points. Data are presented as mean ± SD and normalized to the basal expression of E-cadherin mRNA in nonstimulated MCF-7pl cells (Time zero). (B) Representative Western Blots showing the levels of E-cadherin in IL-1β-stimulated MCF-7pl and MCF-7A3 cells at the times when E-cadherin mRNA expression was evaluated. Protein blots were challenged with anti-E-cadherin antibody. (C) Densitometric analysis of Western Blots against E-cadherin in Triton X-100 soluble and insoluble cell fractions in MCF-7pl and MCF-7A3 cells stimulated with IL-1β obtained from three independent experiments. Data is presented as normalized densitometry relative to time zero, where sum of densitometry values from both Triton X-100 insoluble and soluble E-cadherin bands was set equal to 1. *. (D) Immunolocalization of E-cadherin in MCF-7pl and MCF-7A3 cells. Nonstimulated or IL-1β-stimulated cells were fixed and stained for immunofluorescence with anti-E-cadherin antibody and a secondary antibody tagged with FITC and nuclei counter-stained with DAPI. Bar = 20 μm. Micrographs (e) and (f) are magnifications of the cells within the rectangles in micrographs (b) and (d), respectively. Arrows indicate localization of E-cadherin.
