Establishing Human Gastric Carcinoma Cell Line SGC-7901 With High Expression Of RASSF1A

Objectives: The purpose of the study is to establish human gastric carcinoma cell line SGC-7901 with high expression of RASSF1A so that its roles in apoptosis of tumor cells, cell cycle arrest and cell proliferation inhibition can be further researched.Methods: The technique of RT-PCR was used to amplify full-length cDNA of human gene RASSF1A from gastric tissue, and the full-length cDNA of RASSF1A was inserted into eukaryotic expression vector pcDNA3.1(+) to obtain recombinant plasmid by using technology of DNA recombination. Subsequently, the recombinant plasmid was transfected into SGC-7901 via liposomes. Finally, through screening positive cells clone G418, the expression of RASSF1A was tested by using technology of RT-PCR,Western-bloting.Results: Endonuclease analysis and results of gene sequencing demonstrated RASSF1A had been completely and exactly inserted into pcDNA3.1(+); the best concentration of screening is 500μg/ml in human gastric cancer cell line G418; the results from RT-PCR and Western-blot show expression level increased in transfectd cells.Conclusions: A human gastric cancer cell line which has a stable expression of RASSF1A gene was successfully established by liposomes.