Abstract:

In order to study the role of bone sialoprotein(BSP)in the metastasis of breast cancer cells to bones.a breast cancer cell line stably expressing BSP should be established.In this paper,hbsp gene was subcloned from constructed pB-hBSP vector and was then inserted into pIRES2-EGFP,thus constructing the recombinant plasmid pIRES2-hBSP-EGFP.Moreover.by the transfection of breast cancer cell MDA.MB-231 BO mediated by Lipo-fectamine 2000,the corresponding transfection conditions were optimized to improve the transfection efficiency.Stably transfected breast cancer cells were finally obtained by the selection via G41 8 and flow cytometry.It is found that,in a 96-well cell culture plate,the cell growth confluence is up to 90% ,and that when 1 μg of Lipofecta.mineTM 2000 is used to mediate 280 ng of recombinant plasmid transfecting breast cancer cells for 10 h . the trance.tion eficiency is up to(19.70±1.10)% .

Key words: bone sialoprotein, fluorescent protein, recombinant expression vector, breast cancer cell, transfec-tion, optimization