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Objective To explore the influence of tranSCription factor FOXC2 in angiogenesis of breast cancer MCF-7 cells and to clarify the action mechanism of FOXC2 in promoting tumor angiogenesis.Methods FOXC2 gene and empty vector gene were transfected into breast cancer of MCF-7 cell line with FOXC2 lentivirus gene transfection technique to obtain stable transfection cell line. The MCF-7 cells were devided into non-transfected group,empty-vector group and over-expression group.Matrigel assay and Transwell chamber test were used to observe the changes of tube formation and migration ability of human umbilical vein endothelial cells (HUVECs)in MCF-7 cells supernatant in various groups. PT-PCR and Western blotting methods were applied to detect the expressions of FOXC2,DLL4 and Notch1 mRNA and protein.Results Compared with non-tranfected group and empty-vector group,the tube formation and the migration number of HUVECs in FOXC2 over-expression group were increased(P<0.05);the expressions of FOXC2,DLL4 and Notch1 mRNA and proteins were significantly increased(P<0.05).Conclusion The FOXC2 over-expression in MCF-7 cells can increase the tube formation ability and migration ability of HUVECs,and its mechanism may be related to Notch signaling pathway.
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Objective:In order to study the effect of forkhead box c2 (Foxc2) on the development of skeleton and aorta Methods:Rat monoclonal anti mouse Foxc2 antibody was prepared by using recombinant fusion protein GST Foxc2, and used it to detect the expression of Foxc2 protein in C2C12 myoblasts and embryo of mouse Results:The titer of rat anti mouse Foxc2 antibody was 1: 5 000 This monoclonal antibody can specifically recognized Foxc2 protein producing in mouse L cells and human bladder carcinoma HTB9 cells transfected with CX Foxc2 plasmid Endogenous Foxc2 protein was detected in the mouse myoblast C2C12 cells and increased significantly in C2C12 cells after treatment with bone morphogenetic protein 2(BMP 2) Endogenous Foxc2 protein level was lowered markedly by transfecting C2C12 cells with antisense Foxc2 sequence Foxc2 protein in 11 5 dpc embryo was localized in the developing selerotome and mesenchymal tissue around the aorta Conclusion:Rat anti mouse Foxc2 monoclonal antibody was successfully prepared by using recombinant fusion protein GST Foxc2 The BMP 2 induced Foxc2 protein may play an important role in regulating the osteoblastic differentiation of C2C12 myoblasts and in the formation of axial skeleton and aortic arch