Cross Talk Mechanism among EMT, ROS, and Histone Acetylation in Phorbol Ester-Treated Human Breast Cancer MCF-7 Cells

Figure 3

TPA-elicited Slug induction is regulated by histone H3 acetylation within its promoter region. (a) MCF-7 cells were treated with 1 nM TPA or 1 μM of TSA for the indicated times (for TPA) or 1 h (for TSA). Acetylated histones H3 and H4 were then determined by Western blotting. Values are the mean of fold changes from vehicle-treated cells (). (b) Cells were treated with 1 μM TSA for 6 h, followed by RT-PCR. RT-PCR data were normalized using β-actin levels ( versus vehicle). (c) Cells were treated with 1 nM TPA for 1 h. A ChIP assay was then performed. Relative binding to the promoter region is expressed as the percentage amount over input (%) ( versus vehicle). (d) After cells had been treated with 1 nM TPA for 1 h, HDAC activities were measured. (e) Cells were pretreated with 30 μM garcinol (Gar) or 50 μM CPTH2 (CP) for 30 min and then treated with 1 nM TPA for 6 h, followed by RT-PCR. RT-PCR data were normalized using β-actin levels ( versus vehicle).

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