pEgr-P16重组质粒的构建及其在EC9706细胞中的辐射诱导表达  

CONSTRUCTION OF pEgr-P16 PLASMID AND ITS EXPRESSION IN EC9706 CELL INDUCED BY IONIZING IRRADIATION

作　　者：吴丛梅[1] 黄天华[1] 吴德生[2] 谢庆东[1] 徐小虎[3] 

机构地区：[1]汕头大学医学院生殖医学研究中心,广东汕头515031 [2]兰州医学院 [3]汕头大学医学院法医学教研室,广东汕头515031

出　　处：《癌变．畸变．突变》2003年第4期203-205,共3页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金对外交流与合作项目(No.国科金生外(2002)30210103904号) ;广东省科技计划项目(No.2003C30304)

摘　　要：目的 :构建 pEgr-P16重组质粒并检测其在人食管癌细胞系EC9706中的辐射诱导表达。 方法 :人P16cDNA基因连接到Egr-1启动子的下游构建成 pEgr-P16重组质粒 ,利用脂质体介导转染人EC9706细胞 ,用Westernblot方法检测不同剂量γ射线照射后被转染细胞中P16的表达。结果 :酶切鉴定证实 pEgr-P16重组质粒构建正确。被 pEgr-P16重组质粒转染的人食管癌EC9706细胞经不同剂量γ射线照射后 ,P16基因表达均高于未照射组。 结论Purpose: To construct pEgr-P16 plasmid and detect its expression in esophageal squamous EC9706 cells induced by irradiation. Methods: Human P16 cDNA was ligated to downstream of Egr-1 promoter to construct pEgr-P16 plasmid. The recombined plasmids were transfected into EC9706 cells with liposome. The expression of P16 after different doses of γ-ray irradiation was detected by Westernblot technique. Results: Restriction enzyme digestion showed pEgr-P16 was correctly constructed. The P16 expression in cells transfected with pEgr-P16 induced by different doses of irradiation was higher than that of sham-irradiation group. Conclusion: γ-ray can induce and enhance expression of pEgr-P16 plasmid in EC9706 cells.

关 键 词：EGR-1启动子 Γ射线 P16表达 食管癌细胞系 WESTERNBLOT 

分 类 号：R146[医药卫生—公共卫生与预防医学]