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Compared to both the parental and Ad-GFP controls, FLI1 expression reduced the number of migrating cells by 71% (Figure?3A). The re-expression of FLI1 also reduced the number of invading cells by 87% ( Figure?3B). Therefore, the re-expression of FLI1 significantly reduces the migratory and invasive potential of invasive breast cancer cells. Reciprocal loss-of-function studies were performed by knocking Thymidine kinase down FLI1 in MCF-10A cells using two different shRNA lentiviruses (one targeting the 3�� UTR, shFLI1 #1; the other targets the coding sequence, shFLI1 #2). Verification of knockdown was determined by Western blot analysis (Figure?3C), where shFLI #1 had 70% and shFLI1 #2 had 90% knockdown when compared to parental and short hairpin control. Compared Syk inhibitor to both the parental and shRNA controls, the knockdown of FLI1 expression increased the number of migrating cells by 48% for shFLI1 #1 and 70% for shFLI1 #2 ( Figure?3D). The knockdown of FLI1 expression also increased the number of invading cells by 43% for shFLI1 #1 and 71% for shFLI1 #2 ( Figure?3E). To demonstrate that the change in migration and invasion was the result of knocking down FLI1 and not due to off target effects of the short hairpin, the MCF-10A cells stably infected with shFLI1 #1 were transiently transfected with a plasmid vector expressing FLI1. shFLI1 #1 was selected for the rescue experiment due to its target sequence within the 3��UTR. The re-expression of FLI1 in the MCF-10A shFLI1 #1 cells significantly decreased the number of cells able to migrate and invade compared to MCF-10A shFLI1 #1 cells transfected with Flag control (59% and 70% decrease; respectively) (Figure?3E). Taken together these results suggest that FLI1 contributes to the regulation/of migration and invasion in breast cell lines. Fli1 knockout mice were created by homologous recombination [17], backcrossed to FVB/N mice and then crossed to mice expressing the MMTV-PyVT transgene [32]. The Fli1 homozygous knockout is embryonic lethal so heterozygous knockout mice were compared to wild-type FVB/N mice. Median tumor-free survival was shorter for the MMTV-PyVT-Fli1 heterozygotes (59.5 days) than the wild type (70 days, P selleck inhibitor also grew faster (rate of growth +/? 34.4 mm3/day vs. wt. 26.8 mm3/day; P