Esophageal Cancer
Copyright ©The Author(s) 2004. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Oct 15, 2004; 10(20): 2927-2930
Published online Oct 15, 2004. doi: 10.3748/wjg.v10.i20.2927
Construction of pETNF-P16 plasmid and its expression properties in EC9706 cell line induced by X-ray irradiation
Cong-Mei Wu, Tian-Hua Huang, Qing-Dong Xie, De-Sheng Wu, Xiao-Hu Xu
Cong-Mei Wu, Tian-Hua Huang, Qing-Dong Xie, Research Center of Reproductive Medicine, Shantou University Medical College, Shantou 515041, Guangdong Province, China
Xiao-Hu Xu, Department of Forensic Medicine, Shantou University Medical College, Shantou 515041, Guangdong Province, China
De-Sheng Wu, Lanzhou Medical College, Lanzhou 730000, Gansu Province, China
Author contributions: All authors contributed equally to the work.
Supported by the National Natural Science Foundation of China, No. 30210103904 and the Science and Technology Program of Guangdong Province, No.2003C30304
Correspondence to: Dr. Tian-Hua Huang, Research Center of Reproductive Medicine, Shantou University Medical College, Shantou 515041, Guangdong Province, China. thhuang@stu.edu.cn Telephone: +86-754-8900442 Fax: +86-754-8557562
Received: November 17, 2003
Revised: November 22, 2003
Accepted: December 8, 2003
Published online: October 15, 2004
Abstract

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. TNFα expressions were significantly increased in the transfected cells after different doses of X-ray irradiation than in those after 0Gy irradiation (1192.330-2026.518 pg/mL, P < 0.05-0.01), and the TNFα 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 TNFα 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.

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