Inhibitory effects of silencing S100A4 gene by shRNA on the growth and invasiveness of breast cancer MCF-7 cells.

Objective:To study the inhibitory effects of short hairpin RNA(shRNA) targeting S100A4 gene on the growth and invasiveness of breast cancer MCF-7 cells.Methods:The S100A4 specific shRNA expression vector was constructed and confirmed by sequencing analysis.S100A4-shRNA expression vector was transfected into MCF-7cells via lipofectamineTM 2000,and the expression levels of S100A4 mRNA and protein were determined by real-time fluorogentic quantitative PCR and Western blotting at 48 h after transfection.Flow cytometry and MTT assay were performed to assess the effect of the S100A4-shRNA on the apoptosis and proliferation of MCF-7 cells.The cell line with stable expression of S100A4 was obtained by G418 screening and colony culture.Invasion and migration capability of stably transfected MCF-7 cells was evaluated by using Transwell chamber model and wound healing assay in vitro.The proliferation of stably transfected MCF-7 cells was evaluated by tumor formation test in nude mice.Results: S100A4-shRNA expression vector was successfully constructed and transfected into MCF-7 cells.At 48 h after transfection,S100A4 mRNA and protein expression levels were decreased significantly and the proliferation of MCF-7 cells was markedly suppressed.Transfection of S100A4-shRNA expression vector induced significant apoptosis of MCF-7 cells.The morphology of stably transfected MCF-7 cells did not change,but the invasion and migration capability decreased significantly.Tumor formation test indicated that the tumor size and weight were greatly decreased compared with control group and negative group.Conclusion:S100A4-shRNA expression vector effectively suppresses S100A4 expression in breast cancer MCF-7 cells,and thereby decreasing the proliferation,invasion,and migration capability of MCF-7 cells.