Construction Of Human Carcinoembryonic Antigen Gene Vaccination And Its Expression In Human Tongue Cancer Cell Line In Vitro

OBJECTIVE: (1) To construct human carcinoembryonic antigen (CEA) gene vaccination. (2)To dectect the expression of CEA in human tongue cancer cell line Tca8113.METHODS: (1)Human CEA gene was amplified by RT–PCR method from human colon cancer cells and constructed into eukaryotic expression vector pcDNA3.1-CEA. (2) Human CEA gene in the vector pcDNA3.1-CEA was identified by enzyme digestion , PCR amplification and DNA sequencing. (3) Tca8113 cells were transfected with human pcDNA3.1-CEA as experimental group and eukaryotic expression vector as experimental control respectively, while the non-transfected cells as negative control. The expression of human CEA was detected by RT–PCR and immunohistochemical analysis respectively after the transfection .RESULTS: (1)The cloned DNA was confirmed to be human CEA gene by enzyme digestion , PCR amplification and DNA sequencing. (2) Compared with experimental and negative controls, the expression of CEA mRNA and protein were significantly increased in the pcDNA3.1-CEA group (P<0.05); there was no significantly statistical difference between experimental control and negative one about the CEA expression (p>0.05).CONCLUSION: CEA gene vaccination was successfully constructed and expressed in human tongue squamous cell carcinoma cell line Tca8113 after transfection .