DJ-1真核表达载体构建及其转染口腔癌细胞的表达  

Construction of DJ-1 eukaryotic expression vetor and identification of transfected oral cancer cell line

作　　者：钟永荣[1] 谢思明[2] 程燕飞[3] 殷操[1] 

机构地区：[1]广东省口腔医院,广东广州510280 [2]暨南大学口腔医学系,广东广州510632 [3]广东省中医院口腔科,广东广州510120

出　　处：《暨南大学学报（自然科学与医学版）》2013年第4期401-404,共4页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广东省科技计划项目(2011B080701054)

摘　　要：目的:构建人DJ-1真核表达载体pcDNA3.1(+)/DJ-1,并转染人口腔癌细胞系Tca8113,建立含有DJ-1真核表达载体的口腔癌细胞模型。方法:以人HEK293细胞为模板,应用反转录聚合酶链反应(RT-PCR)法扩增人DJ-1基因全长,与真核表达载体pcDNA3.1(+)重组,经酶切鉴定和测序分析后,通过脂质体介导法转染人口腔癌细胞系Tca8113,并设立空质粒对照组(转染pcDNA3.1空载体)和未转染组,最后采用RT-PCR法检测Tca8113细胞中DJ-1基因的表达情况。结果:RT-PCR法获得长度为500 bp左右的阳性产物,重组质粒后经酶切鉴定和序列分析证实真核表达载体pcDNA3.1(+)/DJ-1构建成功。转染Tca8113细胞后RT-PCR法证实与空质粒组和未转染组相比,转染真核表达载体pcDNA3.1(+)/DJ-1的细胞系中DJ-1基因表达明显增高。结论:成功构建人真核表达载体pcDNA3.1(+)/DJ-1,并建立含有该载体的口腔癌细胞模型,为进一步研究DJ-1基因在口腔癌中的功能及应用DJ-1进行基因治疗奠定基础。Aim：To construct human DJ-1 eukaryotic expression vector pcDNA3.1 （ ＋ ）/DJ-1 and transfect into oral cancer cell line Tca8113, in order to establishment an oral cancer cell model containing DJ-1 expression vector. Methods： RT-PCR method was carried out to clone DJ-1 gene using the mRNA from human HEK293 cell line. Mterwards, DJ-1 gene was recombinated with pcDNA3.1 （ ＋ ） vector and identificated by restriction enzyme digestion and DNA sequencing technology. This recombinated vector was transfected into oral cancer cell line Tca8113 mediated by lipofectamine 2000, with pcDA3.1 （ ＋ ） vector as a blank control. The expression of DJ-1 gene in these cell lines was detected by RT-PCR method. Results： By RT-PCR method, a positve product about 500bp was obtained. The recombinated vector of pcDNA3.1 （ ＋ ） and DJ-1 gene was identified by restriction enzyme digestion and DNA sequencing technology, confirming that eukaryotic expression vector pcDNA3.1 （ ＋ ）/D J-1 was constructed successfully. After transfected into oral cancer cell line Tca8113, the expression level of DJ-1 was significantly

关 键 词：DJ-1基因 载体构建 口腔癌 

分 类 号：R780.2[医药卫生—口腔医学]