【摘要】 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。更多还原

【Abstract】 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.更多还原