癌变·畸变·突变 ›› 2003, Vol. 15 ›› Issue (4): 203-204.doi: 10.3969/j.issn.1004-616x.2003.04.004

• 论著 • 上一篇    下一篇

pEgr-P16重组质粒的构建及其在EC9706细胞中的辐射诱导表达

吴丛梅1,黄天华1,吴德生,谢庆东1 ,徐小虎2   

  1. 1.汕头大学医学院生殖医学研究中心,广东 汕头 515031;2.汕头大学医学院法医学教研室,广东 汕头 515031
  • 收稿日期:2003-06-30 修回日期:2003-07-16 出版日期:2003-10-30 发布日期:2003-10-30
  • 通讯作者: 黄天华

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

WU Cong-Mei1, HUANG Tian-Hua1,WU De-Sheng, et al   

  1. 1. Research Center of Reproductive Medicine, Shantou University Medica College, Shantou 515031, China; 2. Department of forensic medicine, Shantou University Medical College, Shantou 515031, China
  • Received:2003-06-30 Revised:2003-07-16 Online:2003-10-30 Published:2003-10-30
  • Contact: HUANG Tian-Hua

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P16 cDNA基因连接到Egr-1启动子的下游构建成pEgr-P 16重组质粒,利用脂质体介导转染人EC9706细胞,用Western blot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-P 16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:γ射线可诱导pEgr-P16重组质粒在人EC9706细胞中表达增强。

关键词: Egr-1启动子, γ射线, P 16表达, 食管癌细胞系, Western blot

Abstract: 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.

Key words: EGR-1 promoter, γ-ray, P16 expression, Western blot, esophageal squamous cell line