【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EC970 6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EC970 6细胞。利用ELISA ,Westernblot和免疫细胞化学的方法检测pETNF P16在被转染的EC970 6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EC970 6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 0 5或P <0 0 1)。 2GyX射线照射后 2～ 4 8hTNFα和P16的表达量明显高于对照组 (P <0 0 5或P <0 0 0 1)。在正常的EC970 6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EC970 6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更多还原

【Abstract】 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.\;更多还原