人精子蛋白17放射诱导表达细胞模型的构建

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。

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

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 pEgr-Sp17 plasmid in HO8910 cells,the hSp17 radiation-inducible expression ovarian cancer cell model has been constructed successfully.