pEgr-MDA7重组质粒辐射诱导特性的检测

The Irradiation-Induced Property of pEgr-MDA7 Recombinant Plasmid

背景与目的克隆人MDA7cDNA基因,构建pEgr-MDA7重组质粒并检测其在人食管癌细胞系EC9706中经辐射诱导后mRNA水平。材料与方法从人外周血mRNA中克隆人MDA7cDNA基因,连接到Egr-1启动子的下游构建成pEgr-MDA7重组质粒,利用脂质体介导转染人EC9706细胞,用定量PCR方法检测不同剂量X射线照射后被转染细胞中MDA7的mRNA水平。结果测序结果证实MDA7cDNA基因序列正确,酶切鉴定证实pEgr-MDA7重组质粒构建正确。被pEgr-MDA7重组质粒转染的人食管癌EC9706细胞经不同剂量X射线照射后,MDA7基因mRNA水平均高于未照射组。结论X射线可诱导pEgr-MDA7重组质粒在人EC9706细胞中表达增强。

BACKGROUND ＆ AIM： To clone human MDA7 cDNA gene, construct pEgr-MDA7 plasmid, and measure its mRNA level in esophageal squamous EC9706 cells induced by irradiation. MATERIAL AND METHODS： Human MDA7 cDNA gene was isolated from peripheral blood and was then ligated to downstream of Egr-1 promoter to construct pEgr-MDA7 plasmid. The recombinant plasmids were transfected into EC9706 cells with liposome, then MDA7 mRNA level of after different doses of X-ray irradiation was quantified by PCR. RESULTS： Sequencing and restriction enzyme digestion were performed ensuring that MDA7 cDNA gene cloning and construction of pEgr-MDA7 were correct. MDA7 mRNA level in cells transfected with pEgr-MDA7 induced by different doses of irradiation was higher than that of sham-irradiation group. CONCLUSION： X-ray could induce and enhance expression of pEgr-MDA7 gene in EC9706 cells.