pEgr-MDA7重组质粒辐射诱导特性的检测

doi:10.3969/j.issn.1004-616x.2006.05.013

癌变·畸变·突变 ›› 2006, Vol. 18 ›› Issue (5): 381-383.doi: 10.3969/j.issn.1004-616x.2006.05.013

pEgr-MDA7重组质粒辐射诱导特性的检测

吴丛梅1;黄天华1;吴德生1;谢庆东1;徐小虎2

1．汕头大学医学院生殖医学研究中心，广东汕头 515041；2．汕头大学法医学教研室，广东汕头 515041

收稿日期:2005-01-28 修回日期:2005-04-20 出版日期:2006-09-30 发布日期:2006-09-30
通讯作者: 吴丛梅

The Irradiation-Induced Property of pEgr-MDA7 Recombinant Plasmid

WU Cong-mei1, HUANG Tian-hua1, Wu De-sheng1,XIE Qing-dong1,XU Xiao-hu2

1.Research Center of Reproductive Medicine,Shantou University Medical College, Shantou 515041, Guangdong,China;2.Department of Forensic Medicine,Shantou University Medical College,Shantou 515041,Guang Dong,China

Received:2005-01-28 Revised:2005-04-20 Online:2006-09-30 Published:2006-09-30
Contact: WU Cong-mei

摘要/Abstract

摘要： 背景与目的：克隆人MDA7 cDNA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MDA7 cDNA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MDA7 cDNA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。

关键词: MDA-7基因, pEgr-MDA7重组质粒, 辐射

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.

Key words: melanoma differentiation associated gene, pEgr-MDA7 recombinant plasmid, irradiation

吴丛梅, 黄天华, 吴德生, 谢庆东, 徐小虎. pEgr-MDA7重组质粒辐射诱导特性的检测[J]. 癌变·畸变·突变, 2006, 18(5): 381-383.

WU Cong-mei, HUANG Tian-hua, Wu De-sheng, XIE Qing-dong, XU Xiao-hu. The Irradiation-Induced Property of pEgr-MDA7 Recombinant Plasmid[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2006, 18(5): 381-383.

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pEgr-MDA7重组质粒辐射诱导特性的检测

The Irradiation-Induced Property of pEgr-MDA7 Recombinant Plasmid

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