Abstract:

Objective    To construct the pEGFP-N1-ZIP10 expression vector and observe its expression in human breast cancer MCF-7 and MDA-MB-231 cell lines, and also to detect expressions of other zinc transporters when ZIP10 is over-expressed. Methods    The target sequence of ZIP10 was obtained and amplified from human blood by RT-PCR. Then, the cDNA segment was cloned into eukaryote plasmid pEGFP-N1. The pEGFP-N1-ZIP10 was identified by restriction enzyme digestion and checked by DNA sequence analysis. MCF-7 and MDA-MB-231 cells were transiently transfected, and expressions of ZIP10 and other zinc transporters were detected by RT-PCR. Results    Identification of pEGFP-N1-ZIP10 by enzyme digestion and PCR showed that the length, location of insertion and direction of the target gene inserted into the recombinant were correct. After the transfection, over-expression of ZIP10 was found in human breast cancer MCF-7 and MDA-MB-231 cells, while expression of ZIP1 was significantly reduced in the transfected cells. Conclusion     The eukaryotic expression plasmid pEGFP-N1-ZIP10 has been successfully constructed and it can be expressed transiently in MCF-7 and  MDA-MB-231 cells.  The decreased expression of ZIP1 in the transfected cells may indicate the role of ZIP10 in breast carcinogenesis and development.

Key words: Breast cancer cells; ZIP10; ZIP1; Expression vector