小鼠MIP-3α基因在Lewis肺癌中的放射诱导表达

Expression of MIP-3α gene regulated by irradiation via Egr-1 promotor in Lewis lung cancer

目的 构建pEgr 1 MIP 3α(pEM)质粒 ,探讨辐射对被转染的Lewis肺癌细胞中MIP 3α表达的影响。方法 用定向克隆方法 ,构建pEM重组表达质粒 ,脂质体介导转染Lewis肺癌细胞 ,用RT PCR和ELISA方法检测不同剂量电子线照射后的MIP 3α表达水平。结果 本实验构建的重组质粒pEM中MIP 3αcDNA正确插入表达载体 ;各照射组在不同剂量电子线照射后 ,MIP 3α表达均高于未转染组和假照射组。结论 Egr 1启动子具有辐射启动和诱导下游MIP 3α基因在Lewis肺癌中增强表达的功能 ,为pEM用于放射 -基因治疗的体内研究 ,提供了实验基础。

Objective To construct the expression vector of early growth response-1 promoter-macrophage inflammatory protein 3α (pEgr-1-MIP-3α, pEM) to study MIP-3α expression in transfected Lewis lung cancer cells. Methods MIP-3α cDNA was inserted into plasmid with Egr-1 promoter by directional cloning. The vectors were transfected into Lewis lung cancer cells with liposomes. The expression of MIP-3α in transfected Lewis lung cancer cells induced by electron ray irradiation at different doses were confirmed by RT-PCR and Western blotting. Results The recombinant vector was sequenced, and the result indicated the correct insertion in the expression vector. After electron ray irradiation at different doses, MIP-3α expression in the transfected Lewis lung cancer cells increased significantly. Conclusion Electron ray can enhance the expression of MIP-3α in Lewis lung cancer cells. This has laid the experimental foundation for the studies of pEM for radio-genetic therapy in vivo.