| 吴丛梅,黄天华,谢庆东,李德锐,吴德生,徐小虎.pETNF-P16质粒的构建及其在食管癌细胞中的辐射诱导表达特性[J].中华放射医学与防护杂志,2004,24(5):406-408 |
| pETNF-P16质粒的构建及其在食管癌细胞中的辐射诱导表达特性 |
| Construction of pETNF-P16 plasmid and its expression properties in esophageal carcinoma cells induced by ionizing irradiation |
| 投稿时间：2003-10-04 |
| DOI： |
| 中文关键词: Egr-1启动子 X射线 TNFα P16 食管癌 |
| 英文关键词:Egr-1 promoter X-ray TNFα P16 Esophageal carcinoma |
| 基金项目:国家自然科学基金资助项目(30210103904);广东省科技计划资助项目(2003C30304) |
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| 中文摘要: |
| 目的 构建重组质粒pETNF-P16并研究其在食管鳞状细胞癌细胞系EC9706中X射线照射诱导表达特性和基因放射治疗食管癌的可行性。方法 构建重组质粒pETNF-P16,通过脂质体介导转染EC9706细胞。利用ELISA,Western blot和免疫细胞化学的方法检测pETNF-P16在被转染的EC9706细胞中的X射线照射诱导表达特性。结果 成功构建真核载体pETNF-P16并转染EC9706细胞。在不同剂量X射线照射后被转染细胞中TNFα的表达量明显高于0Gy组(P<0.05或P<0.01)。2GyX射线照射后2～48hTNFα和P16的表达量明显高于对照组(P<0.05或P<0.001)。在正常的EC9706细胞中没有P16的表达,在转染组和辐射诱导组中P16有较强的表达。结论 在pETNF-P16质粒转染的EC9706细胞中X射线可诱导TNFα和P16的表达增强。本研究结果为临床食管癌基因放射治疗进一步研究提供实验基础。 |
| 英文摘要: |
| Objective The recombined plasmid pETNF-P16 was constructed to investigate its expression properties in esophageal squamous cell carcinoma cell line EC9706 induced by X-ray irradiation and the feasibility of gene-radiotherapy for esophageal carcinoma. Methods The 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 The eukaryotic expression vector pETNF-P16 was successfully constructed and transfected into EC9706 cells.The expressions of TNFα were significantly increased in the transfected cells after different dose of X-ray irradiation than that of 0 Gy group(P<0.05～0.01) and the expressions of TNFα and P16 were significantly higher during 2～48 h after 2 Gy X-ray irradiation than that of control group (P<0.05～0.01).No P16 expression in normal EC9706 cells and strong expression in the transfected group and irradiation induction group were detected. Conclusions X-ray irradiation induction could significantly enhance the TNFα and P16 expression property in the EC9706 cells transfected with pETNF-P16 plasmid.These results may establish important experimental basis for gene-radiotherapy of esophageal carcinoma. |
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