The effect of specific inhibition of Semaphorin4C (Sema4C) gene expression by small interfering RNA on the malignant behavior of human breast cancer cell line MDA-MB-231

Abstract

Objective: To study the effect of specific inhibition of Semaphorin 4C (Sema4C) gene expression by small interfering RNA (siRNA) on the migration, invasion and proliferation of highly metastatic human breast cancer cell line MDA-MB-231, in vitro. Methods: The Sema4C siRNA was designed by web based software, synthesized and then transfected into MDA-MB-231 cell line by lipofectamine 2000 reagent. Western blot was used to analyze the effect of the siRNA transfection. The migration, invasion and proliferation of the Sema4C-siRNA transfected cells were assessed by monolayer wound healing assay, transwell cell invasion assay and CFSE-flow cytometry. The expression of p-AKT protein in untransfected cells, negative control-transfected MDA-MB-231 cells and Sema4C-siRNA-transfected MDA-MB-231 cells were studied by Western blot analysis. Results: Significant down-regulation of the Sema4C protein was found at 72 hours after transfection of the Sema4C-siRNA. Compared with those of untransfected cells and negative control transfected cells, the migration, invasion and proliferation ability of Sema4C-siRNA-transfected MDA-MB-231 cells were notably weakened. The protein level of p-AKT was lower in the Sema4C-siRNA-transfected MDA-MB-231 cells. Conclusion: The in vitro transient transfection of synthesized Sema4C-siRNA on the MDA-MB-231 cell line can down-regulate the expression of Sema4C protein in the cells. Sema4C-siRNA plays a vital role in suppressing migration, invasion and proliferation of MDA-MB-231 cells, which may be related to PI3K-AKT cell signal pathway.