Abstract:

Objective
 To constructed an eukaryotic expression vector pcDNA3.1(+)-AKR1B10 and  observe its expression in breast cancer MCF-7 cells,and to provide experimental basis for  study on the relationship between AKR1B10 gene and breast cancer.Methods The  breast cancer  MCF-7 cells were transfected transiently with  the constructed eukaryotic expression vector pcDNA3.1(+)-AKR1B10. The expressions of AKR1B10 gene in the untransfected MCF-7 cells,the MCF-7 cells transfected with pcDNA3.1(+)-AKR1B10 and the MCF-7 cells transfected with pcDNA3.1(+) (control vector) were detected by RT-PCR and Western blotting methods.Results The eukaryotic expression vector pcDNA3.1(+)-AKR1B10 was successfully constructed.The RT-PCR results showed that if the AKR1B10 gene expression in the untransfected  MCF-7 cells was regarded as 1, the expression values  were 1.79 in the cells  transfected with pcDNA3.1(+)-AKR1B10 and  0.98 in  the cells tranfected with pcDNA3.1(+). The expression of AKR1B10 gene in the cells  transfected with pcDNA3.1(+)-AKR1B10 was significantly increased compared with the cells   transfected with pcDNA3.1(+) and the untransfected cells (P<0.05).The Western blotting results showed that the expression of AKR1B10 gene in the cells transfected with pcDNA3.1(+)-AKR1B10  was significantly increased compared with the cells transfected with pcDNA3.1(+)  and the untransfected cells.Conclusion The eukaryotic expression vector pcDNA3.1-AKR1B10 is successfully constructed.The AKR1B10 gene highly expresses in the MCF-7 cells transfected with the expression vectror.

Key words: AKR1B10, overexpression vector, transfection