由Egr-1调控TK基因灭活胰腺癌细胞的实验研究  

The killing effect on pancreatic cancer cell in vitro by the radio-inducible TK suicide gene that controlled by Egr-1 promoter

作　　者：刘金龙[1] 钱清[1] 刘训良[1] 郭治源[1] 杜青[1] 郭仕英[2] 李朝军[2] 

机构地区：[1]南京医科大学第一附属医院普外科,江苏南京210029 [2]南京师范大学生命科学学院分子医学实验室,江苏南京210097

出　　处：《南京医科大学学报（自然科学版）》2007年第11期1244-1247,共4页Journal of Nanjing Medical University(Natural Sciences)

摘　　要：目的:观察对放射敏感的早期生长反应基因-1(early growth response-1,Egr-l)启动子调控单纯疱疹病毒胸苷激酶(herpes simplex virus thymidine kinase,TK)基因是否对胰腺癌细胞的杀伤作用有增敏效应。方法:以细菌内同源重组法构建含TK基因的腺病毒载体pAdEgr-1-TK,转染人胰腺癌细胞株PC-3,60Co-γ射线照射后,逆转录-聚合酶链反应(RT-PCR)半定量分析各不同照射剂量组TK基因的mRNA的表达。加入前药丙氧鸟苷(ganciclovir,GCV),MTT法检测其对胰腺癌细胞的杀伤作用。结果:经60Co-γ射线照射后,与对照组胰腺癌细胞(0.86±0.11)相比,前药GCV可明显提高对转染pAdEgr-1-TK胰腺癌细胞的杀伤效率(0.08±0.03)(P<0.001)。结论:由Egr-l启动子调控的TK自杀基因在γ射线作用下可以显著提高杀伤胰腺癌细胞的能力。Objective：To observe the killing effect on pancreatic carcinoma （PC-3） cell lines by early growth response-1 （Egr-1） promoter activating herpes simplex virus thymidine kinase （TK）. Methods ：Adenoviral vector of pAdEgr-l-TK was generated through homologous recombination in bacteria and was transfected to human PC-3.Afier exposured to γ-radiation by a ^60Co souree,hemi-quantitated by RT-PCR respectively to detect the expression of mRNA of TK in different radiation dose groups, in which 0 Gy act as control group. Then the cells were added prodrug ganciclovir （GCV）,the survival rate was evaluated by MTr method. Results：After irradiation,transfected cell lines （0.08 ± 0.03） were killed by prodrug GCV at higher percentage significantly compared with control group （0.86 ± 0.11 ）（P 〈 0.001 ）. Conclusion：It indicated that the Egr-1 promoter caused high expression of TK gene in cancer cells after exposured to ^60Co-γ,which should significantly boost the killing effect on pancreatic cancer cell with the prodrug.

关 键 词：胰腺癌 Γ射线 基因治疗 EGR-1启动子 

分 类 号：R735.9[医药卫生—肿瘤]