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


CONCLUSIONS
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.
Wu
Cong-Mei
CM
Research Center of Reproductive Medicine, Shantou University Medical College, Shantou 515041, Guangdong Province, China.
Huang
Tian-Hua
TH
Xie
Qing-Dong
QD
Wu
De-Sheng
DS
Xu
Xiao-Hu
XH
D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000532:radiotherapy*; D003131:Combined Modality Therapy; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000532:radiotherapy*; D015870:Gene Expression; D015316:Genetic Therapy; D006801:Humans; D010957:Plasmids / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism*; D014162:Transfection / Q000379:methods; D014407:Tumor Cells, Cultured / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha