摘要
目的 利用可被放射线激活而转录的早期生长反应基因 1 (Egr 1 )启动子驱动单纯疱疹病毒胸苷激酶基因 (tk)在肝癌细胞中高效表达。方法 构建以Egr 1为启动子 ,以tk为目的基因的重组质粒 pET ,经脂质体介导转染人肝癌细胞株SMMC 772 1 ,经G41 8抗性筛选、分别以 0、5、7.5、1 0、1 5、2 0Gy剂量γ射线照射、逆转录 聚合酶链反应 (RT PCR)半定量分析、比较tk基因的mRNA表达。结果 转染后的肝癌细胞株经射线照射后其tkmRNA的表达较未照射组 (55 .2±7.2 )有明显提高 ,以 1 5Gy最为显著 (1 1 7.2± 1 1 .1 ,P <0 .0 0 1 )。结论 经γ射线照射后 ,可被放射线转录激活的Egr 1启动子可引起tk基因在肝癌细胞中高效表达 ,从而为肝癌的基因
Objective To improve the efficacy of herpes simplex virus thymidine kinase (tk) expression by the way of radiation inducible early growth response 1 (Egr 1) promoter.Methods Plasmid pET was constructed by fusing of Egr 1 promoter to the upstream of tk gene and transfect human hepatocellular carcinoma (HCC) cell line (SMMC 7721) with lipofectamine as a delivery system. The cloned cells had been selected with G418. After exposure to γ radiation by a 60 Co source, cellular tk mRNA expression in hepatoma cells was compared by RT PCR analysis.Results After irradiation, the tk mRNA expression of transfected cell lines was increased markedly compare with non irradiation cell lines (55.2±7.2), especially in 15?Gy dosage (117.2±11.1, P<0.001).Conclusions As a radio inducible promoter, Egr 1 can activates tk expression significantly in transduced hepatoma cell line after irradiation. These in vitro data will provide an experimental basis for gene therapy and for further research of gene radiotherapy in HCC.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2002年第5期419-420,共2页
Chinese Journal of Experimental Surgery
关键词
Γ射线
启动子
肝细胞癌
基因疗法
治疗
Gamma rays
Promoter
Hepatocellular carcinoma
Gene therapy
