人精子蛋白17放射诱导表达细胞模型的构建  

Construction of Human Sperm Protein 17 Radiation-Inducible Expression Ovarian Cancer Cell Model

作　　者：张春华[1] 李芳秋[1] 杨爱龙[1] 缪家文[1] 朱锡旭[2] 

机构地区：[1]南京军区南京总医院全军医学检验中心,江苏南京210002 [2]南京军区南京总医院放射治疗科,江苏南京210002

出　　处：《生物技术通讯》2007年第3期454-456,共3页Letters in Biotechnology

基　　金：国家自然科学基金项目(30670599);南京市科技发展计划项目(200401070-5);江苏省"六大人才高峰"基金项目(2005A2)

摘　　要：目的:构建pEgr-Sp17重组质粒,并检测其在人卵巢癌细胞系HO8910中的辐射诱导表达,探索Egr-1基因调控序列辐射诱导下游基因表达的功能,建立放射诱导表达人精子蛋白Sp17的细胞模型。方法:用双酶切、粘端连接的方法构建含有辐射诱导特性的早期反应因子Egr-1和人Sp17基因的pEgr-Sp17重组质粒,利用脂质体介导其转染人HO8910细胞,用免疫细胞化学方法检测X线照射后被转染细胞中Sp17的表达。在确证Sp17能瞬时表达后,继续用含G418的RPMI1640培养基培养细胞,筛选稳定表达Sp17的细胞株。结果:测序结果和酶切鉴定均证实pEgr-Sp17重组质粒构建正确;免疫细胞化学试验表明pEgr-Sp17重组质粒转染的人HO8910细胞表达Sp17蛋白。结论:X线可诱导pEgr-Sp17重组质粒在人HO8910细胞中表达Sp17蛋白,构建了Sp17蛋白放射诱导表达的卵巢癌细胞模型。Objective： To investigate the expression of sperm protein 17（Sp17） induced by radiation in ovarian cancer cell line HO8910, explore whether Egr-1 promoter could induce expression of gene downstream and construct radiation-inducible expression ovarian cancer cell model. Methods： p-Egr-Sp17 vector was constructed by restriction enzyme digestion and ligation, and was introduced into HO8910 cells by LipofectAMINE. The expression of Sp17 after X-ray irradiation was detected by immunocytochemistry technique. The HO8910 cells expressing hSp17 stably were obtained through G418 screening. Results： Sequencing and restriction enzyme digestion showed pEgr-Sp17 was correctly constructed. The HO8910 cells transfected recombinant plasmid showed Sp17 expression. Conclusion： Radiation could induce expression of pEgrSp17 plasmid in HO8910 cells, the hSp17 radiation-inducible expression ovarian cancer cell model has been constructed successfully.

关 键 词：精子蛋白Sp17 EGR-1启动子 X线 表达 卵巢癌细胞模型 

分 类 号：Q78[生物学—分子生物学] Q813