Vascular Endothelial Growth Factor Receptor-1Activation Induces Epithelial-to Mesenchymal Transition In Breast Carcinoma Cell Line MCF-7

OBJECTIVE： Comparing the alteration of E-cadherin， Snail and Vimentinexpressing in the breast carcinoma cell line MCF-7with the vascular endothelial growthfactor receptor-1（VEGFR-1） before and after MCF-7activation via treatment with theVEGFR-1specific ligand VEGF-B. To explore the relationship between the activation ofVEGFR-1and epithelial-to-mesenchymal transition （EMT）in breast carcinomaMCF-7cell line from the molecule and protein levels.Targeting VEGFR-1may providethe theory foundation for the development of therapeutic approaches for this disease.METHODS:Through the culture of breast carcinoma cell line MCF-7expressingthe receptor VEGFR-1in vitro, MTT method was used to find the specific VEGFR-1ligand VEGFB to promote breast cancer cell line MCF-7growth optimumconcentration；breast carcinoma cell line was cultured with this concentration so that theVEGFR-1of MCF-7was activated.To investigate the changes about the expression ofE-cadherin、Snail、Vimentin mRNA and protein before and after VEGFR-1activationin breast carcinoma cell line MCF-7via real-time fluorescence quantitative PCR（FQ-PCR）and Western-blot methodRESULTS：After VEGFR-1activation，E-cadherin、Vimentin and Snail proteinexpression were0.399±0.02、0.919±0.17、0.460±0.012respectively；before activation，expression were0.824±0.41、0.207±0.067、0.134±0.054respectively by Western-blot.E-cadherin expression level was decreased obviously，on the contrary，Vimentin and Snailprotein expression level was increased significantly after VEGFR-1activation（P<0.05）.E-cadherin、Vimentin、Snail gene mean Ct（cycle threshold value，Ct）were29.45、12.45、15.205respectively after VEGFR-1activation；were12.785、25.55、20.55respectively before VEGFR-1activation.The result of2-ΔΔCtshow the gene copy ratio before and after activation were0.1811、103.97、40.645respectively. ΔCt value was-9.565±0.02、3.2±0.05、-1.8±0.05and7.1±0.10、-9.9±0.05、-7.145±0.005before and afteractivation respectively. statistically significant differences were observed by thecomparison of E-cadherin、Vimentin、Snail gene expression.（P<0.05，α=0.05）。CONCLUSION：The study demonstrate,from protein and gene level,VEGF-B caninduce EMT in breast cancer cell line MCF-7expressing the receptor VEGFR-1；VEGF-B/VEGFR-1signaling pathway may provide a new target for breast cancer genetheraphy.