Figure 4
Motility and proapoptotic effects of testosterone-HSA in the absence or presence of inhibitors in Caco2 cells. (A) Cells both in and out of the presence of 10−6 mol/L anastrozole or 10−6 mol/L flutamide were cultured with 10−7 mol/L testosterone-HSA on the Transwell culture chambers, provided with an 8-µm pore-size polycarbonate filter, according to the manufacturer’s instructions. After 24 h, invaded cells, attached to the lower surface of the filter, were stained with DAPI. The slices were imaged under the microscope, and the number of cells in 10 random fields was counted. Bars represent the % of cell invasion in control and treated cells; (n = 3), ***P < 0.001. (B) Quantitative APOP-ercentage apoptosis assay of TAC-stimulated Caco2 cells and similar experiments in the presence of anastrozole. Cells were exposed or not exposed to 10−7 mol/L testosterone-HSA for 24 h and proapoptotic responses were assessed by the APOPercentage apoptosis assay. Equally, cells pretreated or not pretreated with 10−6 mol/L anastrozole were exposed to testosterone-HSA for 24 h. Cells serum starved for comparable periods of time served as a positive control for apoptosis. Bars present the mean OD measured at 550 nm; *** P < 0.001, n = 3.