Transfer of siRNA against CD73 suppresses human breast cancer cell line MB-MDA-231 adhesion to extracellular matrix / 中国病理生理杂志

ABSTRACT

AIM:To study the effect of CD73 on breast cancer cell line MB-MDA-231 adhesion to extracellular matrix.METHODS: ① CD73 siRNA plasmid was constructed and transfected into MB-MDA-231 cells by lipofectamine 2000.② The transfection efficiency was analyzed by flow cytometry using eGFP as a marker gene.Stable transfected MB-MDA-231 cells were selected using G418.③ RT-PCR and Western blotting analysis of CD73 expression in MB-MDA-231 cells was performed.④ The effects of CD73 on MB-MDA-231 cells adhesion to extracellular matrix were assessed by cell adhesion assay.RESULTS: ① Transfection of MB-MDA-231 cells with CD73 siRNA plasmid significantly inhibited CD73 expression at mRNA level.The efficiency was up to 91%.② Transfection of MB-MDA-231 cells with CD73 siRNA plasmid significantly inhibited CD73 expression at a protein level.The efficiency was up to 79.3%.③ Treatment of MB-MDA-231 cells with CD73 siRNA resulted in diminished adhesion to extracellular matrix.CONCLUSION: This study demonstrates that CD73 siRNA effectively inhibits CD73 gene expression in MB-MDA-231 cells leading to adhesion to extracellular matrix suppression.