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Construction of pETNF-P16 plasmid and its expression properties in EC9706 cell line induced by X-ray irradiation
Cong-Mei Wu1, Tian-Hua Huang, Qing-Dong Xie
1Research Center of Reproductive Medicine, Shantou University Medical College, Shantou 515041, Guangdong Province, China.
Aim:
Recombined plasmid pETNF-P16 was constructed to investigate its expression properties in esophageal squamous carcinoma cell line EC9706 induced by X-ray irradiation and the feasibility of gene-radiotherapy for esophageal carcinoma.
Methods:
Recombined plasmid pETNF-P16 was constructed and transfected into EC9706 cells with lipofectamine. ELISA, Western blot, and immunocytochemistry were performed to determine the expression properties of pETNF-P16 in EC9706 after transfection induced by X-ray irradiation.
Results:
Eukaryotic expression vector pETNF-P16 was successfully constructed and transfected into EC9706 cells. TNFalpha expressions were significantly increased in the transfected cells after different doses of X-ray irradiation than in those after 0Gy irradiation (1,192.330-2,026.518 pg/mL, P<0.05-0.01), and the TNFalpha expressions and P16 were significantly higher 6-48 h after 2 Gy X-ray irradiation (358.963-585.571 pg/mL, P<0.05-0.001). No P16 expression was detected in normal EC9706 cells. However, there was strong expression in the transfected and irradiation groups.
Conclusion:
X-ray irradiation induction could significantly enhance TNFalpha and P16 expression in EC9706 cells transfected with pETNF-P16 plasmid. These results may provide important experimental data and therapeutic potential for gene-radiotherapy of esophageal carcinoma.
