FIG. 4.

Young ASC secretome enhances proliferation and estrogen signaling in MCF-7 cell lines. MCF-7 cells were treated with condition media from aged or young ASCs, and proliferation using crystal violet was evaluated at days 0, 1, 5, and 7. MCF-7 cells treated with young CM showed greater proliferation through (A) visual staining and (B) absorption. N = 3 biological and technical replicates, error bars = SEM (C) MCF-7 cells were cultured with aged and young ASC CM and collected for qRT-PCR. Both PGR and SDF-1 expression were significantly different (*P < 0.05) from aged expression. N = 3 biological and N = 2 technical replicates, error bars = SEM (D) MCF-7 cells were cultured with aged and young ASC CM and evaluated for total and phosphorylated AKT, ERK, and ER-α protein expression using WES capillary system. Results show an increase at p-ER ser-167 in MCF-7 cells stimulated with young CM. (E) MCF-7 cells were treated with tamoxifen for 4 h before being treated with either young or aged ASC CM. MCF-7 cells were collected after 24 h for qRT-PCR to evaluate ER regulated gene expression. Results confirm that ASC signaling is mediated by ER-α. N = 3 biological and N = 2 technical replicates, error bars = SEM. ERK, extracellular signal related kinase; IGF-1, insulin like growth factor 1; PGR, progesterone receptor.