Construction of pETNF-P16 plasmid and its expression properties in EC9706 cell line induced by X-ray irradiation

AIM:Recombined plasmid pETNF-P16 was constructed toinvestigate its expression properties in esophagealsquamous carcinoma cell line EC9706 induced by X-rayirradiation and the feasibility of gene-radiotherapy foresophageal carcinoma.METHODS:Recombined plasmid pETNF-P16 was constructedand transfected into EC9706 cells with lipofectamine.ELISA,Western blot,and immunocytochemistry were performedto determine the expression properties of pETNF-P16 inEC9706 after transfection induced by X-ray irradiation.RESULTS:Eukaryotic expression vector pETNF-P16 wassuccessfully constructed and transfected into EC9706 cells.TNFα expressions were significantly increased in thetransfected cells after different doses of X-ray irradiation thanin those after 0Gy irradiation(1 192.330-2 026.518 pg/mL,P<0.05-0.01),and the TNFα expressions and P16 weresignificantly higher 6-48 h after 2 Gy X-ray irradiation(358.963-585.571 pg/mL,P<0.05-0.001).No P16 expressionwas detected in normal EC9706 cells.However,there wasstrong expression in the transfected and irradiation groups.CONCLUSION:X-ray irradiation induction could significantlyenhance TNFα and P16 expression in EC9706 cellstransfected with pETNF-P16 plasmid.These results mayprovide important experimental data and therapeuticpotential for gene-radiotherapy of esophageal carcinoma.