(B) Colony formation assay was used to measure cell proliferative capacity in MDA-MB-231 cells treated with control siRNA or BIRC5 siRNA. *, P < 0.05. Repressing LASP1 expression could inhibit migration of MDA-MB-231 cells To investigate the effect of LASP1 on selleck chemical the migration of TNBC cell, we evaluated the cell migratory capacity of MDA-MB-231 cells transfected with LASP1 siRNA (or control siRNA). The expression of LASP1 protein in the cells transfected with LASP1 siRNA was significantly decreased in comparison with that of cells transfected with control siRNA (Figure 5A), indicating that the expression of LASP1 was effectively inhibited by LASP1 siRNA. Subsequent studies
showed that the migratory capacity of cells transfected with LASP1 siRNA was significantly lower than that of cells treated with control siRNA (Figure 5B). Figure 5 Repressing LASP1 expression could
VX-680 inhibit migration of MDA-MB-231 cells. (A) Immunoblots of LASP1 protein in MDA-MB-231 cells treated with control siRNA or LASP1 siRNA. β-actin was used as a TSA HDAC price loading control. (B) Transwell migration assay was performed to detect the migratory capacity of MDA-MB-231 cells treated with control siRNA or LASP1 siRNA. *, P < 0.05. The inhibition of MDA-MB-231 cell proliferation by miR-203 is attenuated by the over-expression of BIRC5 To provide direct evidence that down-regulation of BIRC5 is required for the anti-tumorigenic effects of miR-203, we transfected MDA-MB-231 cells with pcDNA-BIRC5 and miR-203 precursor. We first confirmed that BIRC5 and miR-203 have been conducted into the cells (Figure 6A), then, we used colony formation assay to show that the inhibition of MDA-MB-231 cell proliferation by miR-203 could be partially rescued by BIRC5 up-regulated (Figure 6B). These data clearly indicate that the ectopic over-expression of BIRC5 could efficiently block the effect on proliferation caused by miR-203. Figure 6 Over-expression of BIRC5 could significantly attenuate the effect of miR-203 on the inhibition of MDA-MB-231 cell
proliferation. (A) BIRC5 protein expression was detected by western blot and normalized to β-actin protein ADP ribosylation factor levels. (B) Colony formation assay was performed to detect proliferative capacity in MDA-MB-231 cells. *, P < 0.05. The inhibition of MDA-MB-231 cell migration by miR-203 is attenuated by the over-expression of LASP1 To provide direct evidence that miR-203 inhibits the migration of TNBC cells through the LASP1-mediated signal pathway, we transfected MDA-MB-231 cells with miR-203 precursor and pcDNA-LASP1. We confirmed the effect of the transfection by western blot (Figure 7A). The migration assay showed that the over-expression of LASP1 could partially rescue the migratory capacity of MDA-MB-231 cells treated with the miR-203 precursor (Figure 7B). Figure 7 Over-expression of LASP1 significantly attenuated the effect of miR-203 on the inhibition of MDA-MB-231 cell migration.