Figure 5. Anti-TNF-α treatment effect on pro-motility activity and mammosphers (MS) promotion of HUVECs secretome on MCF-7 tumor cell.

A. Drawing showing the experimental design. Senescent cells exposed to anti-TNF-α long-term treatment and senescent control cells were switched to fresh medium for 24 h and then their conditioned medium was mixed with 1/3 fresh DMEM. These mixtures, senescence conditioned medium (SEN-CM), and anti-TNF-α treated senescence conditioned medium (ATT SEN-CM) were used to treat MCF-7 in a wound healing assay. Conditioned medium from young cells was mixed with DMEM and used as control. B. Light microscopic photographs showing MCF-7 migration in the wound healing assay 0, 4 and 8 h after treatment with SEN-CM or ATT SEN-CM. C. Percentage of migrating cells after 4 and 8 h exposure to SEN-CM and ATT SEN-CM. Data expressed as percent of control. D. Representative pictures of MCF7-derived mammosphers (MS) promotion induced by conditioned media obtained from young cells (CTRL-CM), senescent cells (SEN-CM), and anti-TNF-α treated senescent cells (ATT SEN-CM). E. Quantification of MCF7-derived mammospheres in presence of different conditioned media. Data are presented as number of MS per well, at 3 and 7 days. Data are mean ± S.D. of 3 independent experiments. Scale bar 50 μm. * p < 0.05. * Student's t test, p < 0.05. Data are mean ± S.D. of 3 independent experiments.