Establishment of a cell line of Tca8113 with stable expression of nm23-H1 by gene transfection / 实用口腔医学杂志

ABSTRACT

Objective:To establish a tongue cancer cell line that stably overexpresses nm23-H1.Methods:The reconstructed plasmid, pCMV-BamH1-Neo-nm23-H1, was transfected into bacteria and amplified. The plasmid was identified by electrophoresis on a 10 g/L agarose gel and stained with ethidium bromide and then transfected to the cell line of Tca8113 by lipofectin strategy and was selected by G418 for 5 weeks. The stable expression of nm23-H1 was identified by immunofluorescence,Western-blotting and flow cytometer.Results:Restriction endonuclease Bam H1 examination showed that 986 bp of nm23-H1 was in pCMV-Bam-Neo vector. 5 weeks after transfection positive clones were obtained and the transfected cells were proliferated to confluent.Immunohistochemical examination,Western blot and flow cytometry showed that nm23-H1 expression was increased in the transfected cells.Conclusion:A tongue cancer cell line expressing nm23-H1 is established.