The establishment and identification of the stable transfected SMMC7721 cell expressing NNMT.

Objective To construct a stable transfected of human hepatoma cancer cell line SMMC7721 that can highly express human NNMT.Method NNMT gene was amplified from recombinant plasmid pGEX4T-1 /NNMT by PCR method,and cloned into vector pMD19-T.Then cloned into eukaryotic expression vector pcDNA3.1(-)and the recombinant plasmid was named as pcDNA3.1-NNMT.Lipofectamine 2000 was used to transfect the pcDNA3.1-NNMT into SMMC7721 cells.The NNMT-transfected SMMC7721 cells were selected with G418 for 4 weeks.Finally,RT-PCR was used to detect the expression of human NNMT in SMMC7721 cells.Result We have successfully constructed the eukaryotic expression vector for the expression of NNMT.Conclusion This study indicated NNMT gene could be stably expressed in SMMC7721 cells and its functions on tumor cells could be further analyzed by this stable cell line.