Transduction Of HOXB7 Gene Impact On Invasion Of SKBR-3 Breast Cancer Cell Line

Purpose We use the Coding sequence(CDS)of HOXB-7 gene sense transducted into breast carcinoma cell line SKBR-3 to enhance its expression and to investigate its effect on cell invasiveness of breast carcinoma cell line SKBR-3.Methods Clone and amplified (PCR method) HOXB-7 gene from MCF-7 cell line.Then construct an expression plasmid containing HOXB-7 gene. Two human breast cancer cell lines MCF-7,SKBR-3 were analyzed for the expression of H0XB7 mRNA,bFGF mRNA by RT-PCR and their protein by Western blot.SKBR-3 cells were divided into 3 groups:untreated control group,no-specific group transfected with PIRES2-EGFP(negtive), and HOXB-7 group transfected with recombined plasmid PIRES2-H0XB7. SKBR-3 cells were stably transfected with PIRES2-EGFP and PIRES2-HOXB7. RT-PCR and Western blot were used to examine the HOXB7,bFGF mRNA and protein expression.The invasion of cells transfected with PIRES2-EGFP and PIRES2-HOXB7 or non-transfected cells was observed through matrigel invasion assay,gelatin zymography and Western blotting.Results Only MCF-7 cells expressed H0XB7 and bFGF, non-transfected SKBR-3 cells did not;The PIRES2-HOXB7 transfected SKBR-3 cells expressed HOXB7 and bFGF . The motility and invasion of SKBR-3 cells stably expressing PIRES2-HOXB7 were increased compared with cells stably expressing PIRES2-EGFP or non-transfected cells(P< 0.01).MMP-2 protein and mRNA were barely detected in both group,but MMP-9 mRNA can be detected in the PIRES2-HOXB7 transfected group.Conclusions SKBR-3 cells did not express HOXB7 gene and bFGF. But they were detectable in SKBR-3 cells after sense transfected with plasmid PIRES2-HOXB7 and effectively increase the invasion of the SKBR-3 cell,which suggested that HOXB7 plays an important role in invasion of breast cancer.